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Immunology Cell Type Markers CD Non-lineage
使用敲除细胞株进行验证重组RabMAb

重组Anti-PD-L1抗体[28-8] (ab205921)

  • Datasheet
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Reviews (15)Q&A (1)References (303)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunocytochemistry - Anti-PD-L1 antibody [28-8] (ab205921)
  • Flow Cytometry - Anti-PD-L1 antibody [28-8] (ab205921)
  • Western blot - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunocytochemistry - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Frozen sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Western blot - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Western blot - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
  • Anti-PD-L1 antibody [28-8] (ab205921)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [28-8] to PD-L1
  • Suitable for: ICC, IHC-P, WB, Flow Cyt, IHC-Fr
  • Knockout validated
  • Reacts with: Human, Recombinant fragment

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 APC Carrier Free FITC HRP PE

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概述

  • 产品名称

    Anti-PD-L1抗体[28-8]
    参阅全部 PD-L1 一抗
  • 描述

    兔单克隆抗体[28-8] to PD-L1
  • 宿主

    Rabbit
  • 经测试应用

    适用于: ICC, IHC-P, WB, Flow Cyt, IHC-Frmore details
  • 种属反应性

    与反应: Human, Recombinant fragment
  • 免疫原

    Recombinant full length protein corresponding to Human PD-L1 (extracellular). The immunogen contains the specific extracellular domain of huPD-L1 (Phe19-Thr239). See reference for more info - www.ncbi.nlm.nih.gov/pmc/articles/PMC4561627/
    Database link: Q9NZQ7

  • 阳性对照

    • Tissue: Human tonsil, head and neck squamous cell carcinoma and placenta tissues; L2987 cell line. Cell Lines: Positives: B-CPAP (high), ES-2 (medium), HCC70 (low), CHO-PDL1, U-87 MG For additional information - please refer to this publication: Programmed death-ligand 1 (PD-L1) expression in various tumor types - http://www.immunotherapyofcancer.org/content/1/S1/P53 IHC-Fr: Frozen human tonsil tissue sections
  • 常规说明

    FURTHER INFORMATION ON POSITIVE CONTROLS (Chinese version)

    Tissue:
    - Tonsil - with hyperreactive changes. Screening of hyper-reactive tonsils is recommended to find tonsil with the highest expression of PD-L1 in crypt epithelium, macrophages homing the germinal centers and interfollicular mononuclear leukocytes.
    - Tumor tissues - prescreened for positive tumor and inflammatory infiltrates. PD-L1 expression varies by tumor type so screening is recommended to find positive and negative tumor controls.
    - The following publication is useful for finding suitable tumor types for PD-L1 expression: 
    http://www.immunotherapyofcancer.org/content/1/S1/P53
    - Note: Look for specimens with high numbers of inflammatory macrophages and mononuclear leukocytes.

    Cell Lines:
    - Positive controls: B-CPAP (high), ES-2 (medium), HCC70 (low).

    Primary antibody negative control: Recombinant Rabbit IgG isotype control antibody, ab172730.

    Recombinant protein positive control: Recombinant human PD-L1 protein, ab167713.

    Immunohistochemistry usage:
    For IHC usage on FFPE tissues, we recommend using PD-L1 IHC panel ab236676, which contains PD-L1 antibody clone 28-8 (ab205921), HIER antigen retrieval reagent (ab208572) and IHC detection kit HRP/DAB (ab209101).

    Western blot usage:
    For clone 28-8, it is recommended to use Odyssey system. This system has the advantages of a wider dynamic range and less background than chemiluminescence.

    This PD-L1 antibody [28-8] has been used as detector antibody in Human PD-L1 SimpleStep ELISA® kit: ab214565 and ab277712.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    28-8
  • 同种型

    IgG
  • 研究领域

    • Immunology
    • Cell Type Markers
    • CD
    • Non-lineage
    • Immunology
    • Immune System Diseases
    • Autoimmune
    • Cancer
    • Tumor immunology
    • Tumor-associated antigens
    • Cancer
    • Tumor biomarkers
    • Tumor antigens
    • Signal Transduction
    • Antibodies
    • pd-l1

相关产品

  • Alternative Versions

    • APC Anti-PD-L1 antibody [28-8] (ab206967)
    • Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)
    • Alexa Fluor® 488 Anti-PD-L1 antibody [28-8] (ab209959)
    • Alexa Fluor® 647 Anti-PD-L1 antibody [28-8] - Extracellular domain (ab209960)
    • HRP Anti-PD-L1 antibody [28-8] (ab209961)
    • PE Anti-PD-L1 antibody [28-8] (ab209962)
    • Alexa Fluor® 555 Anti-PD-L1 antibody [28-8] (ab213358)
    • Alexa Fluor® 568 Anti-PD-L1 antibody [28-8] (ab213359)
    • Alexa Fluor® 594 Anti-PD-L1 antibody [28-8] (ab213360)
    • FITC Anti-PD-L1 antibody [28-8] (ab224027)
    • Anti-PD-L1 antibody [28-8] - BSA and Azide free (ab228413)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Immunohistochemistry kits

    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human PD-L1 protein (Active) (ab167713)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Anti-PD1 antibody [EPR4877(2)] (ab137132)
    • Anti-PD1 antibody [NAT105] (ab234444)
    • Anti-PD1 antibody [NAT105] (ab52587)
  • Unmasking reagent

    • Universal HIER antigen retrieval reagent (10X) (ab208572)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab205921于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC
Use a concentration of 2 µg/ml.
IHC-P (7)
Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Please refer to protocol section here.

For antigen buffer for FFPE tissue, it is recommended to use Universal HIER antigen retrieval reagent (ab208572).

WB (5)
Use at an assay dependent concentration. Detects a band of approximately 40-60 kDa (predicted molecular weight: 33 kDa).

Please refer to protocol section here.

Flow Cyt
Use at an assay dependent concentration.

Please refer to protocol section here.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr (1)
Use a concentration of 1 µg/ml.
说明
ICC
Use a concentration of 2 µg/ml.
IHC-P
Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Please refer to protocol section here.

For antigen buffer for FFPE tissue, it is recommended to use Universal HIER antigen retrieval reagent (ab208572).

WB
Use at an assay dependent concentration. Detects a band of approximately 40-60 kDa (predicted molecular weight: 33 kDa).

Please refer to protocol section here.

Flow Cyt
Use at an assay dependent concentration.

Please refer to protocol section here.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr
Use a concentration of 1 µg/ml.

靶标

  • 功能

    Involved in the costimulatory signal, essential for T-cell proliferation and production of IL10 and IFNG, in an IL2-dependent and a PDCD1-independent manner. Interaction with PDCD1 inhibits T-cell proliferation and cytokine production.
  • 组织特异性

    Highly expressed in the heart, skeletal muscle, placenta and lung. Weakly expressed in the thymus, spleen, kidney and liver. Expressed on activated T- and B-cells, dendritic cells, keratinocytes and monocytes.
  • 序列相似性

    Belongs to the immunoglobulin superfamily. BTN/MOG family.
    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • 细胞定位

    Cell membrane and Endomembrane system.
  • Target information above from: UniProt accession Q9NZQ7 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 29126 Human
    • Omim: 605402 Human
    • SwissProt: Q9NZQ7 Human
    • Unigene: 521989 Human
    • 别名

      • B7 H antibody
      • B7 H1 antibody
      • B7 homolog 1 antibody
      • B7-H1 antibody
      • B7H antibody
      • B7H1 antibody
      • CD 274 antibody
      • CD-274 antibody
      • CD274 antibody
      • CD274 antigen antibody
      • CD274 molecule antibody
      • MGC142294 antibody
      • MGC142296 antibody
      • OTTHUMP00000021029 antibody
      • PD L1 antibody
      • PD-L1 antibody
      • PD1L1_HUMAN antibody
      • PDCD1 ligand 1 antibody
      • PDCD1L1 antibody
      • PDCD1LG1 antibody
      • PDL 1 antibody
      • PDL1 antibody
      • Programmed cell death 1 ligand 1 antibody
      • Programmed death ligand 1 antibody
      • RGD1566211 antibody
      see all

    图片

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      IHC image of ab205921 staining PD-L1 in human tonsil formalin fixed paraffin embedded tissue sections*, performed on a Leica BOND RX (Polymer Refine kit). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH9, epitope retrieval solution 2) for 30 mins at 98°C. The section was then incubated with ab205921, 5μg/ml working concentration, for 60 mins at room temperature and detected using an HRP conjugated compact polymer system for 8 minutes at room temperature. DAB was used as the chromogen for 10 minutes at room temperature. The section was then counterstained with hematoxylin, blued, dehydrated, cleared and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Immunocytochemistry - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunocytochemistry - Anti-PD-L1 antibody [28-8] (ab205921)

      Immunocytochemistry analysis of CHO-PDL1 (PD-L1 stably expessed Chinese hamster ovary epithelial cell) labeling PD-L1 with purified ab205921 at 1/400 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.32 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
      Negative controls: Cells not transfected with PD-L1, and both the transfected and mock tranfected cells without the primary antibody.

    • Flow Cytometry - Anti-PD-L1 antibody [28-8] (ab205921)
      Flow Cytometry - Anti-PD-L1 antibody [28-8] (ab205921)

      ab205921 specificity testing by Flow Cytometry (KO testing): Loss of detection on KO cells.
      Strong detection with anti-PD-L1 (ab205921, clone 28-8) TALEN constructs targeting exon4 of human PD-L1, transcript variant 1 (NM_014143.3) and complete knock out (K.O) confirmed by deep sequencing in clone L2-14. Cell surface staining is almost completely eliminated in the L2987 L2-14 KO cell line.

      For recommended Flow Cytometry (Flow Cyt) protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

      Alexa Fluor® 488 (ab209959) and Alexa Fluor® 647 (ab209960) conjugated versions are available for this clone.

    • Western blot - Anti-PD-L1 antibody [28-8] (ab205921)
      Western blot - Anti-PD-L1 antibody [28-8] (ab205921)

      Primary ab Dilution 1:100 dilution, Secondary ab Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody, 1:20,000 dilution, Blocking and diluting buffer and concentration 5% NFDM/TBST, Lane 1: NCI-H1975 (Human non-small cell lung cancer epithelia), Observed MW 40-60 kDa.

      For recommended Western Blot (WB) protocol for endogenous PD-L1 expression, please refer to the protocol book in the protocol section and/or here (downloadable copy).

    • Immunocytochemistry - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunocytochemistry - Anti-PD-L1 antibody [28-8] (ab205921)This image is courtesy of an Abreview submitted by Dr. Dimitra Kalamida.

      Paraformaldehyde-fixed, Triton X-100 permeabilized U-87 MG (human glioblastoma-astrocytoma epithelial cell line) cells stained for PD-L1 (red) using ab205921 at 1/200 dilution in ICC/IF, followed by CF568 Donkey anti-rabbit IgG(H+L) secondary antibody at 1/500 dilution.

      Alexa Fluor® 488 (ab209959) and Alexa Fluor® 647 (ab209960) conjugated versions are available for this clone.

      See Abreview

    • Immunohistochemistry (Frozen sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Frozen sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      IHC image of PD-L1 staining in a section of frozen normal human tonsil* performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab205921, 1ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.


      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.


      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Western blot - Anti-PD-L1 antibody [28-8] (ab205921)
      Western blot - Anti-PD-L1 antibody [28-8] (ab205921)
      All lanes : Anti-PD-L1 antibody [28-8] (ab205921) at 1/100 dilution

      Lane 1 : H1975 (Human non-small cell lung cancer epithelial cell) whole cell lysate
      Lane 2 : NCI-H1299 (Human lung carcinoma epithelial cell) whole cell lysate
      Lane 3 : A549 (Human lung carcinoma epithelial cell) whole cell lysate
      Lane 4 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate
      Lane 5 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
      Lane 6 : SK-BR-3 (Human breast adenocarcinoma epithelial cell) whole cell lysate
      Lane 7 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
      Lane 8 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate
      Lane 9 : PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate
      Lane 10 : DU 145 (Human prostate carcinoma epithelial cell) whole cell lysate
      Lane 11 : A375 (Human malignant melanoma epithelial cell) whole cell lysate
      Lane 12 : MeWo (Human malignant melanoma fibroblast) whole cell lysate
      Lane 13 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate
      Lane 14 : Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
      Lane 15 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
      Lane 16 : BXPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate
      Lane 17 : PANC-1 (Human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate
      Lane 18 : NIH:OVCAR-3 (Human ovary adenocarcinoma epithelial cell) whole cell lysate
      Lane 19 : SK-OV-3 (Human ovarian cancer epithelial cell) whole cell lysate
      Lane 20 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 33 kDa
      Observed band size: 40-60 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Diluting buffer and concentration: 5% NFDM/TBST. Expression of PD-L1 varied widely among the tumor cell lines.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)Nakamura et al PLoS One. 2017; 12(10): e0186192. Published online 2017 Oct 19. doi: 10.1371/journal.pone.0186192

      Formalin-fixed, paraffin-embedded human lung cancer tissue stained for PD-L1 using ab205921.

      Representative images of PD-L1 expression.
      (A) <1.0%, (B) 1.0–4.9%, (C) 5.0–9.9%, (D) 10.0–49.9%, and (E) ≥50.0% PD-L1-positive cells (magnification, 200×).

      From image 3 of Nakamura et al.

      Nakamura et al PLoS One. 2017; 12(10): e0186192. Published online 2017 Oct 19. doi: 10.1371/journal.pone.0186192

      |Reproduced under Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      IHC image of ab205921 staining PD-L1 in PD-L1 Dynamic Range Analyte Control formalin fixed paraffin embedded cell lines (HistoCyte Laboratories), performed on a Leica BOND RX (Polymer Refine kit). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH9, epitope retrieval solution 2) for 30 mins at 98°C. The section was then incubated with ab205921, 5μg/ml working concentration, for 60 mins at room temperature and detected using an HRP conjugated compact polymer system for 8 minutes at room temperature. DAB was used as the chromogen for 10 minutes at room temperature. The section was then counterstained with hematoxylin, blued, dehydrated, cleared and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling PD-L1 with ab205921 at a dilution of 1:400. Heat mediated antigen retrieval was performed using AR9 antigen retrieval solution, and microwave treatment for 15 min at 20% power. Anti-Rabbit/Mouse HRP polymer (PerkinElmer Opal Polymer HRP Ms Plus Rb) was used as secondary antibody. Opal tyramide amplification was performed using Opal 520 fluorophore. Counterstained with DAPI stain. Image scanned with Vectra 3.0 and analyzed via Phenochart software.
      This image was courteously provided by Dr. Houssein Abdul Sater, Georgia Cancer Center.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      Ab205921 specificity testing by Immunohistochemistry (KO testing): Loss of detection on KO Cells

      Strong IHC detection with anti-PD-L1 (ab205921, clone 28-8) is seen in human lung adenocarcinoma tumor cell line L2987. PDL1 gene was edited in L2987 cells using TALEN constructs targeting exon4 of human PD-L1, transcript variant 1 (NM_014143.3) and complete knock out (K.O) confirmed by deep sequencing in clone L2-14. IHC detection is completely eliminated in the L2987 L2-14 K.O. cell line.

       

      For recommended Immunohistochemistry (IHC) protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      Immunohistochemical staining of PD-L1 in formalin fixed, paraffin embedded human non-squamous non-small cell lung cancer (NSQ-NSCLC) using ab205921 at a dilution of 1/400, incubated for an hour at room temperature. Heat mediated antigen retrieval was carried out in low pH buffer and the sample was blocked with peroxidase blocking buffer for 3 minutes.

      This image was courteously provided by Dr. Kai Schmitt from the Institute of Pathology, Saarbrücken-Rastpfuhl.

      See Abreview

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      Immunohistochemical analysis of CHO PD-L1 cells with ab205921 at 2 µg/ml.
      High power view
      A) Rabbit IgG, 5 µg/mL. No staining
      B) Anti PD-L1, 2 µg/mL (ab205921 batches 1)
      C) Anti PD-L1, 2 µg/mL (ab205921 batches 3)
      D) Anti PD-L1, 2 µg/mL (ab205921 batches 4)
      E) Anti PD-L1, 2 µg/mL (ab205921 batches 5)
      F) Anti PD-L1, 2 µg/mL (ab205921 batches 6)
      G) Anti PD-L1, 2 µg/mL (ab205921 batches 7)

       

      All batches/lots (1,3,4,5,6,7) showed consistent results.


      Note strong, moderate, and weak (red, yellow, and white arrows respectively) plasma membrane staining of CHO PD-L1 transfected cells

       

      For recommended Immunohistochemistry (IHC) protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      Immunohistochemical analysis of CHO Parental cells with ab205921 at 2 µg/ml.
      High power view
      A) Rabbit IgG, 5 µg/mL. No staining
      B) Anti PD-L1, 2 µg/mL (ab205921 batches 1)
      C) Anti PD-L1, 2 µg/mL (ab205921 batches 3)
      D) Anti PD-L1, 2 µg/mL (ab205921 batches 4)
      E) Anti PD-L1, 2 µg/mL (ab205921 batches 5)
      F) Anti PD-L1, 2 µg/mL (ab205921 batches 6)
      G) Anti PD-L1, 2 µg/mL (ab205921 batches 7)

       

      All batches/lots (1,3,4,5,6,7) showed consistent results.

       

      Note absence of PD-L1 expression in CHO parental cells.

       

      For recommended Immunohistochemistry (IHC) protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      Immunohistochemical analysis of Human Lung NSCLC with ab205921 at 2 µg/ml.
      High power view
      A) Rabbit IgG, 5 µg/mL. No staining
      B) Anti PD-L1, 2 µg/mL (ab205921 batches 1)
      C) Anti PD-L1, 2 µg/mL (ab205921 batches 3)
      D) Anti PD-L1, 2 µg/mL (ab205921 batches 4)
      E) Anti PD-L1, 2 µg/mL (ab205921 batches 5)
      F) Anti PD-L1, 2 µg/mL (ab205921 batches 6)

       

      All batches/lots (1,3,4,5,6) showed consistent results.


      Note linear and complete or partial (arrows) PD-L1 staining of tumor cells. Tumor associated immune cells localized over the tumor margin exhibit positive plasma membrane staining (small arrows).

       

      For recommended Immunohistochemistry (IHC) protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human non-small cell lung cancer tissue labelling PD-L1 with ab205921. Tumor cells and immuno cells localized within the stroma show PD-LA plasma membrane staining.

      For antigen retrival buffer, Universal HIER antigen retrieval reagent (ab208572) was used.
      For IHC detection kit, Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) is recommended.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human melanoma tissue labelling PD-L1 with ab205921. Tumor cells show weak and partial postive PD-L1 expresseion in the plasma membrane. PD-L1 positive tumor associated immunoe cells are also stained. 

      For antigen retrival buffer, Universal HIER antigen retrieval reagent (ab208572) was used.
      For IHC detection kit, Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) is recommended.

    • Western blot - Anti-PD-L1 antibody [28-8] (ab205921)
      Western blot - Anti-PD-L1 antibody [28-8] (ab205921)

      (A and B) Western blots of recombinant PD-L1 protein (Lane 1), cell lysates of CHO-PD-L1 (Lane 3), CHO (Lane 4), ES-2 (Lane 5) and Colo205 (Lane 6) cell lines. In B, anti-PD-L1 (ab205921, clone 28-8) was pre-incubated with purified recombinant PDL1 protein overnight at 4°C.

      Blank/no sample (Lane2). Lane 2 is blank on purpose.

      For recommended Western Blot (WB) protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human head and neck squamous cell carcinoma tissue labeling PD-L1 with ab205921 at 2 µg/ml. Counterstained with Hematoxylin.

      For antigen retrival buffer, Universal HIER antigen retrieval reagent (ab208572) was used.
      For IHC detection kit, Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) is recommended.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)This image is courtesy of an Abreview submitted by Mr. Rudolf Jung.

      Paraformaldehyde-fixed, paraffin-embedded human placenta tissue stained for PD-L1 using ab205921 at 1/100 dilution in immunohistochemical analysis.

      See Abreview

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [28-8] (ab205921)

      Immunohistochemical analysis of formalin-fixed, paraffin-embedded L2987 (Human lung adenocarcinoma cell line with endogenous PD-L1 expression) cells labeling PD-L1 with ab205921 at 2 µg/ml. Counterstained with Hematoxylin.

      For antigen retrival buffer, Universal HIER antigen retrieval reagent (ab208572) was used.
      For IHC detection kit, Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) is recommended.

    • Anti-PD-L1 antibody [28-8] (ab205921)
      Anti-PD-L1 antibody [28-8] (ab205921)

    实验方案

    • Protocol Booklet
    • Protocol Booklet - General Notes in Chinese

    Click here to view the general protocols

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    文献 (303)

    发表研究结果有使用 ab205921?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab205921 被引用在 303 文献中.

    • Mei J  et al. A comparability study of natural and deglycosylated PD-L1 levels in lung cancer: evidence from immunohistochemical analysis. Mol Cancer 20:11 (2021). IHC-P ; Human . PubMed: 33413365
    • Ozawa N  et al. PD-L1 upregulation is associated with activation of the DNA double-strand break repair pathway in patients with colitic cancer. Sci Rep 11:13077 (2021). IHC-P ; Human . PubMed: 34158547
    • Bolly HMB  et al. Analysis of Mutant Isocitrate Dehydrogenase 1 Immunoexpression, Ki-67 and Programmed Death Ligand 1 in Diffuse Astrocytic Tumours : Study of Single Center in Bandung, Indonesia. J Korean Neurosurg Soc 64:100-109 (2021). PubMed: 33105535
    • Kurose S  et al. Whole-Exome Sequencing of Rare Site Endometriosis-Associated Cancer. Diseases 9:N/A (2021). PubMed: 33557369
    • Wang YL  et al. Expression of PD1/PDL1 in gastric cancer at different microsatellite status and its correlation with infiltrating immune cells in the tumor microenvironment. J Cancer 12:1698-1707 (2021). PubMed: 33613757
    View all Publications for this product

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    1-7 of 7 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PD-L1 antibody [28-8]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (tonsil)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH 6
    Permeabilization
    No
    Specification
    tonsil
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Mar 07 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PD-L1 antibody [28-8]

    Inconclusive
    Abreviews
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Dog Tissue sections (lymph node)
    Antigen retrieval step
    Heat mediated
    Permeabilization
    No
    Specification
    lymph node
    Blocking step
    ultratek kit, ScyTek Laboratories, Logan, Utah, USA as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More

    Sara Belluco

    Verified customer

    提交于 Sep 12 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PD-L1 antibody [28-8]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rabbit Tissue sections (Colon/Tonsil)
    Antigen retrieval step
    Enzymatic - Buffer/Enzyme Used: pH9 20 mins 100oC
    Specification
    Colon/Tonsil
    Blocking step
    No blocking step used for 30 minute(s) · Concentration: 3%
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 May 30 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PD-L1 antibody [28-8]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (NSQ-NSCLC)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EnVision FLEX TRS, Low pH
    Permeabilization
    No
    Specification
    NSQ-NSCLC
    Blocking step
    EnV FLEX Peroxidase-Blocking Reagent as blocking agent for 3 minute(s) · Concentration: 100%
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 May 11 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PD-L1 antibody [28-8]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Placenta)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer pH 9,0
    Permeabilization
    Yes - Commercial Wash Buffer with Tween
    Specification
    Placenta
    Fixative
    Paraformaldehyde
    Read More

    MR. Rudolf Jung

    Verified customer

    提交于 Feb 04 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PD-L1 antibody [28-8]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Placenta)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA pH8.5
    Permeabilization
    No
    Specification
    Placenta
    Blocking step
    Ventana blocking agent as blocking agent for 4 minute(s) · Concentration: 100% · Temperature: 36°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Dec 04 2015

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PD-L1 antibody [28-8]

    Inconclusive
    Abreviews
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Tumor generated by a carcinogen (DMBA) or PANC02 m)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer, pH6
    Permeabilization
    No
    Specification
    Tumor generated by a carcinogen (DMBA) or PANC02 m
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Apr 15 2016

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