重组Anti-HLA-DR抗体[EPR3692] - Low endotoxin,Azide free (ab215985)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3692] to HLA-DR - Low endotoxin, Azide free
- Suitable for: Flow Cyt (Intra), Mass Cytometry, ICC/IF, IHC-P, WB
- Reacts with: Rat, Human
概述
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产品名称
Anti-HLA-DR抗体[EPR3692] - Low endotoxin,Azide free
参阅全部 HLA-DR 一抗 -
描述
兔单克隆抗体[EPR3692] to HLA-DR - Low endotoxin,Azide free -
宿主
Rabbit -
特异性
Signal detected in rat sample is the ortholog of HLA.
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经测试应用
适用于: Flow Cyt (Intra), Mass Cytometry, ICC/IF, IHC-P, WBmore details
不适用于: IP -
种属反应性
与反应: Rat, Human
预测可用于: Recombinant fragment不与反应: Mouse -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HEK-293T cells transfected with a human HLA-DR expression vector containing a his-tag whole cell lysate. Flow Cyt (intra): Raji cells. ICC/IF: Hut-78 cells. IMC: Human tonsil tissue IHC-P: Human tonsil tissue, human skin tissue, human spleen tissue, liver vessels tissue, skin vessels tissue, endometrial carcinoma vessels tissue, human kidney tissue, rat spleen tissue and rat colon tissue.
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常规说明
ab215985 is the carrier-free version of ab92511.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
解离常数(KD)
KD = 1.67 x 10 -10 M Learn more about KD -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR3692 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab215985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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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Mass Cytometry |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 29 kDa.
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说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Mass Cytometry
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 29 kDa. |
靶标
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功能
Binds peptides derived from antigens that access the endocytic route of antigen presenting cells (APC) and presents them on the cell surface for recognition by the CD4 T-cells. The peptide binding cleft accommodates peptides of 10-30 residues. The peptides presented by MHC class II molecules are generated mostly by degradation of proteins that access the endocytic route, where they are processed by lysosomal proteases and other hydrolases. Exogenous antigens that have been endocytosed by the APC are thus readily available for presentation via MHC II molecules, and for this reason this antigen presentation pathway is usually referred to as exogenous. As membrane proteins on their way to degradation in lysosomes as part of their normal turn-over are also contained in the endosomal/lysosomal compartments, exogenous antigens must compete with those derived from endogenous components. Autophagy is also a source of endogenous peptides, autophagosomes constitutively fuse with MHC class II loading compartments. In addition to APCs, other cells of the gastrointestinal tract, such as epithelial cells, express MHC class II molecules and CD74 and act as APCs, which is an unusual trait of the GI tract. To produce a MHC class II molecule that presents an antigen, three MHC class II molecules (heterodimers of an alpha and a beta chain) associate with a CD74 trimer in the ER to form an heterononamer. Soon after the entry of this complex into the endosomal/lysosomal system where antigen processing occurs, CD74 undergoes a sequential degradation by various proteases, including CTSS and CTSL, leaving a small fragment termed CLIP (class-II-associated invariant chain peptide). The removal of CLIP is facilitated by HLA-DM via direct binding to the alpha-beta-CLIP complex so that CLIP is released. HLA-DM stabilizes MHC class II molecules until primary high affinity antigenic peptides are bound. The MHC II molecule bound to a peptide is then transported to the cell membrane surface. In B-cells, the interaction between HLA-DM and MHC class II molecules is regulated by HLA-DO. Primary dendritic cells (DCs) also to express HLA-DO. Lysosomal miroenvironment has been implicated in the regulation of antigen loading into MHC II molecules, increased acidification produces increased proteolysis and efficient peptide loading. -
序列相似性
Belongs to the MHC class II family.
Contains 1 Ig-like C1-type (immunoglobulin-like) domain. -
翻译后修饰
Ubiquitinated by MARCH1 or MARCH8 at Lys-244 leading to down-regulation of MHC class II. When associated with ubiquitination of the beta subunit of HLA-DR: HLA-DRB4 'Lys-254', the down-regulation of MHC class II may be highly effective. -
细胞定位
Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus > trans-Golgi network membrane. Endosome membrane. Lysosome membrane. Late endosome membrane. The MHC class II complex transits through a number of intracellular compartments in the endocytic pathway until it reaches the cell membrane for antigen presentation. - Information by UniProt
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数据库链接
- Entrez Gene: 3122 Human
- Omim: 142860 Human
- SwissProt: P01903 Human
- Unigene: 520048 Human
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别名
- DASS-397D15.1 antibody
- DR alpha chain antibody
- DR alpha chain precursor antibody
see all
图片
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All lanes : Anti-HLA-DR antibody [EPR3692] (ab92511) at 1/1000 dilution
Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with a human HLA-DR expression vector containing a his-tag, whole cell lysate
Lane 2 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with a human HLA-DOA expression vector containing a his-tag, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 29 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking buffer and concentration : 5% NFDM/TBST
Diluting buffer and concentration : 5% NFDM/TBSTThis antibody does not cross-react with human HLA-DOA.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
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ab92511 staining HLA-DR in HuT-78 (human Sezary syndrome) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab7291 and ab150120 were used as counterstains for primary antibody ab92511 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.
Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labelling HLA-DR with ab92511 at 1/10000 dilution (0.07 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) secondary antibody. Positive staining on human tonsil tissue. The section was incubated with ab92511 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control.
Heat mediated antigen retrieval with Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 30 minutes.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
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Overlay histogram showing Raji cells stained with ab92511 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92511, 1/50) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
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Imaging Mass Cytometry™ (IMC™) image of human tonsil tissue stained with Anti-HLA-DR antibody [EPR3692]. ab215985 (carrier-free antibody, purified) was metal-conjugated using a Maxpar® Antibody Labeling Kit from Fluidigm. Immunostaining was performed according to Fluidigm’s protocols. Briefly, slides were subject to deparaffinization and heat-induced epitope retrieval, followed by overnight incubation at 4°C with an antibody cocktail containing metal-tagged antibodies in blocking buffer. Slides were subsequently washed with 0.2% Triton-X and 1x PBS, counterstained with Cell-ID™ Intercalator-Ir diluted at 1/400 in 1x PBS for 30 min at room temperature, rinsed for 5 min with distilled H2O, and air-dried prior to IMC™ acquisition. IMC™ acquisition was performed using the Fluidigm Hyperion™ Imaging System.
Imaging Mass Cytometry™, IMC™, Cell-ID™, Hyperion™ and Maxpar® are trademarks of Fluidigm Canada
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Immunohistochemical analysis of paraffin-embedded human skin tissue labelling HLA-DR with ab92511 at 1/10000 dilution (0.07 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) secondary antibody. Positive staining on human skin tissue. The section was incubated with ab92511 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control.
Heat mediated antigen retrieval with Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 30 minutes.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
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ab92511 staining HLA-DR in human spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/700. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.
Negative control 1: PBS in place of primary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
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ab92511 showing positive staining in Endometrial carcinoma vessels tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
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ab92511, at a 1/100 dilution, staining HLA-DRA in paraffin embedded Human tonsil (A) and Human kidney (B) tissue by Immunohistochemistry. Detection: by DAB staining
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
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Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling HLA DR with ab92511 at a dilution of 1/1000. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody, at a dilution of 1/500. Counter stained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
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Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling HLA DR with ab92511 at a dilution of 1/1000. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody, at a dilution of 1/500. Counter stained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
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Tissue Microarrays stained for "Anti-HLA-DR antibody [EPR3692]” using "ab92511" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab92511 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
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Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (5)
ab215985 被引用在 5 文献中.
- Tong G et al. Intratumoral CD8+ T cells as a potential positive predictor of chemoimmunotherapy response in PD-L1-negative advanced gastric cancer patients: a retrospective cohort study. J Gastrointest Oncol 13:1668-1678 (2022). PubMed: 36092315
- Ferrian S et al. Multiplexed imaging reveals an IFN-γ-driven inflammatory state in nivolumab-associated gastritis. Cell Rep Med 2:100419 (2021). PubMed: 34755133
- Mastropasqua R et al. Corneoscleral limbus in glaucoma patients: in vivo confocal microscopy and immunocytological study. Invest Ophthalmol Vis Sci 56:2050-8 (2015). PubMed: 25744981
- Steain M et al. Analysis of T cell responses during active varicella-zoster virus reactivation in human ganglia. J Virol 88:2704-16 (2014). PubMed: 24352459
- Wang H et al. CD68(+)HLA-DR(+) M1-like macrophages promote motility of HCC cells via NF-?B/FAK pathway. Cancer Lett 345:91-9 (2014). IHC-P ; Human . PubMed: 24333724