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Immunology Adaptive Immunity T Cells CD
使用敲除细胞株进行验证重组RabMAb

重组Anti-CD86抗体[EPR21962] (ab239075)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)
  • Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)
  • Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)
  • Immunoprecipitation - Anti-CD86 antibody [EPR21962] (ab239075)
  • Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
  • Anti-CD86 antibody [EPR21962] (ab239075)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21962] to CD86
  • Suitable for: WB, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free

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概述

  • 产品名称

    Anti-CD86抗体[EPR21962]
    参阅全部 CD86 一抗
  • 描述

    兔单克隆抗体[EPR21962] to CD86
  • 宿主

    Rabbit
  • 经测试应用

    适用于: WB, ICC/IF, Flow Cyt, IPmore details
    不适用于: IHC-P
  • 种属反应性

    与反应: Human
  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: Ramos, Raji and Daudi whole cell lysates. ICC/IF: Raji and Daudi cells. Flow Cyt: Raji cells, Human monocyte-derived dendritic cells.
  • 常规说明

    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: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    EPR21962
  • 同种型

    IgG
  • 研究领域

    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Immunology
    • Cell Type Markers
    • CD
    • Non-lineage
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • B Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Dendritic Cell Lineage
    • Stem Cells
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    • Monocytic Lineage
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • CD markers ELISA kits

相关产品

  • Alternative Versions

    • Anti-CD86 antibody [EPR21962] - BSA and Azide free (ab242020)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • ELISA pair antibody

    • Recombinant human CD86 protein (Fc Chimera) (ab167720)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CD86 knockout Raji cell line (ab273858)
  • KO cell lysates

    • Human CD86 knockout Raji cell lysate (ab273812)
  • Recombinant Protein

    • Recombinant human CD86 protein (Fc Chimera) (ab167720)
  • Related Buffer

    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab239075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
WB
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 38 kDa).
ICC/IF
1/100.
Flow Cyt
1/500.
IP
1/30.
说明
WB
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 38 kDa).
ICC/IF
1/100.
Flow Cyt
1/500.
IP
1/30.
应用说明
Is unsuitable for IHC-P.

靶标

  • 功能

    Receptor involved in the costimulatory signal essential for T-lymphocyte proliferation and interleukin-2 production, by binding CD28 or CTLA-4. May play a critical role in the early events of T-cell activation and costimulation of naive T-cells, such as deciding between immunity and anergy that is made by T-cells within 24 hours after activation. Isoform 2 interferes with the formation of CD86 clusters, and thus acts as a negative regulator of T-cell activation.
  • 组织特异性

    Expressed by activated B-lymphocytes and monocytes.
  • 序列相似性

    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • 翻译后修饰

    Polyubiquitinated; which is promoted by MARCH8 and results in endocytosis and lysosomal degradation.
  • 细胞定位

    Membrane.
  • Target information above from: UniProt accession P42081 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: 942 Human
    • Omim: 601020 Human
    • SwissProt: P42081 Human
    • Unigene: 171182 Human
    • 别名

      • Activation B7-2 antigen 3 antibody
      • Activation B7-2 antigen antibody
      • B-lymphocyte activation antigen B7-2 2 antibody
      • B-lymphocyte activation antigen B7-2 antibody
      • B7 2 antibody
      • B7 antibody
      • B7-2 antibody
      • B7.2 antibody
      • B70 antibody
      • B72 antibody
      • B72 antigen antibody
      • BU63 antibody
      • CD28 antigen ligand 2 2 antibody
      • CD28 antigen ligand 2 antibody
      • Cd28l2 antibody
      • CD28LG2 antibody
      • CD86 antibody
      • CD86 antigen (CD28 antigen ligand 2 B7 2 antigen) antibody
      • CD86 antigen (CD28 antigen ligand 2, B7-2 antigen) 1, 2 antibody
      • CD86 antigen (CD28 antigen ligand 2, B7-2 antigen) antibody
      • CD86 antigen antibody
      • CD86 molecule antibody
      • CD86_HUMAN antibody
      • CLS1 antibody
      • CTLA-4 counter-receptor B7.2 2 antibody
      • CTLA-4 counter-receptor B7.2 2, 3 antibody
      • CTLA-4 counter-receptor B7.2 antibody
      • Early T-cell costimulatory molecule 1 antibody
      • ETC-1 antibody
      • FUN 1 antibody
      • FUN-1 antibody
      • FUN1 antibody
      • LAB72 antibody
      • Ly-58 antibody
      • Ly58 antibody
      • MB7 antibody
      • MB7-2 antibody
      • Membrane glycoprotein antibody
      • MGC34413 antibody
      • T lymphocyte activation antigen CD86 precursor antibody
      • T-lymphocyte activation antigen CD86 antibody
      • TS/A-2 antibody
      see all

    图片

    • Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)
      Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)

      Flow cytometry staining of human monocyte-derived dendritic cells (top) or human monocyte-derived dendritic cells treated with 1μg/mL lipopolysaccharide (LPS) for 24h (bottom), with ab239075 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Monocyte-derived dendritic cells were incubated for 30 min at 4°C in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab239075 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100μl at 0.04 μg/ml (1/56500)) for 30min on ice. The cells were simultaneously stained with CD209.

      The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 4°C

      Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

    • Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)
      Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)

      Flow cytometric analysis of Raji (human Burkitt's lymphoma cell line) cell line labeling CD86 with ab239075 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

      Gated on viable cells.

    • Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)
      Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Daudi (human Burkitt's lymphoma cell line) cells labeling CD86 with ab239075 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Daudi cell line is observed.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
      Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
      All lanes : Anti-CD86 antibody [EPR21962] (ab239075) at 1/1000 dilution

      Lane 1 : Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate
      Lane 2 : CD86 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 38 kDa
      Observed band size: 70 kDa why is the actual band size different from the predicted?



      Lanes 1 - 2: Merged signal (red and green). Green - ab239075 observed at 70 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

      ab239075 was shown to react with CD86 in wild-type Raji cells in western blot with loss of signal observed in CD86 knockout sample. Wild-type and CD86 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab239075 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)
      Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (human Burkitt's lymphoma cell line) cells labeling CD86 with ab239075 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Raji cell line is observed.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunoprecipitation - Anti-CD86 antibody [EPR21962] (ab239075)
      Immunoprecipitation - Anti-CD86 antibody [EPR21962] (ab239075)

      CD86 was immunoprecipitated from 0.35 mg of Raji whole (human Burkitt's lymphoma cell line) cell lysate with ab239075 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab239075 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

      Lane 1: Raji whole cell lysate 10 µg (Input). 

      Lane 2: ab239075 IP in Raji whole lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239075 in Raji whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
      Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
      All lanes : Anti-CD86 antibody [EPR21962] (ab239075) at 1/1000 dilution

      Lane 1 : Ramos (human Burkitt's lymphoma cell line) whole cell lysate
      Lane 2 : Raji (human Burkitt's lymphoma cell line) whole cell lysate
      Lane 3 : Daudi (human Burkitt's lymphoma cell line) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 38 kDa
      Observed band size: 80 kDa why is the actual band size different from the predicted?



      Exposure time : Lane 1: 3 minutes; Lanes 2-3: 26 seconds.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Anti-CD86 antibody [EPR21962] (ab239075)
      Anti-CD86 antibody [EPR21962] (ab239075)

    实验方案

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    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 (21)

    发表研究结果有使用 ab239075?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab239075 被引用在 21 文献中.

    • Zhang M  et al. Exosomal Mir-3613-3p derived from oxygen-glucose deprivation-treated brain microvascular endothelial cell promotes microglial M1 polarization. Cell Mol Biol Lett 28:18 (2023). PubMed: 36870962
    • Hong Y  et al. Microglia-containing cerebral organoids derived from induced pluripotent stem cells for the study of neurological diseases. iScience 26:106267 (2023). PubMed: 36936782
    • Lee CY  et al. Repurposing Astragalus Polysaccharide PG2 for Inhibiting ACE2 and SARS-CoV-2 Spike Syncytial Formation and Anti-Inflammatory Effects. Viruses 15:N/A (2023). PubMed: 36992350
    • Han Z  et al. Ultrasmall iron-quercetin metal natural product nanocomplex with antioxidant and macrophage regulation in rheumatoid arthritis. Acta Pharm Sin B 13:1726-1739 (2023). PubMed: 37139421
    • Liu B  et al. MITA Promotes Macrophage Proinflammatory Polarization and Its circRNA-Related Regulatory Mechanism in Recurrent Miscarriage. Int J Mol Sci 24:N/A (2023). PubMed: 37298501
    View all Publications for this product

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