製品の概要

  • 製品名
    Anti-HLA-DR antibody [TAL 1B5]
    HLA-DR 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [TAL 1B5] to HLA-DR
  • アプリケーション
    適用あり: WB, IHC-P, Flow Cytmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Tissue, cells or virus corresponding to HLA-DR. Bristol 8 separated alpha chain preparation

  • ポジティブ・コントロール
    • Ab20181 gave a positive signal in Raji, Ramos and Daudi whole cell lysates, and in the following human tissue lysates: spleen; tonsil; liver. This antibody gave a positive result in IHC in the following FFPE tissue: Human skin melanoma.
  • 特記事項

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Please note that some batches of ab20181 may contain 0.4M arginine. Please contact Scientific Support for further information.
  • Concentration information loading...
  • 精製度
    Protein G purified
  • ポリ/モノ
    モノクローナル
  • クローン名
    TAL 1B5
  • ミエローマ
    P3-NS1/1-Ag4-1
  • アイソタイプ
    IgG1
  • 軽鎖の種類
    kappa
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab20181 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB Use a concentration of 2 - 5 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 29 kDa).

We recommend using 3% milk as the blocking agent in Western Blot.

IHC-P Use a concentration of 0.1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt Use 0.005-0.01µg for 106 cells.

ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能
    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.
  • 配列類似性
    Belongs to the MHC class II family.
    Contains 1 Ig-like C1-type (immunoglobulin-like) domain.
  • 翻訳後修飾
    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.
  • 細胞内局在
    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
  • 参照データベース
  • 別名
    • DR alpha chain antibody
    • DR alpha chain precursor antibody
    • DRA_HUMAN antibody
    • DRB1 antibody
    • DRB4 antibody
    • Histocompatibility antigen HLA DR alpha antibody
    • HLA class II histocompatibility antigen antibody
    • HLA class II histocompatibility antigen DR alpha chain antibody
    • HLA DR1B antibody
    • HLA DR3B antibody
    • HLA DRA antibody
    • HLA DRA1 antibody
    • HLA DRB1 antibody
    • HLA DRB3 antibody
    • HLA DRB4 antibody
    • HLA DRB5 antibody
    • HLA-DRA antibody
    • HLADR4B antibody
    • HLADRA1 antibody
    • HLADRB antibody
    • Major histocompatibility complex class II DR alpha antibody
    • Major histocompatibility complex class II DR beta 1 antibody
    • Major histocompatibility complex class II DR beta 3 antibody
    • Major histocompatibility complex class II DR beta 4 antibody
    • Major histocompatibility complex class II DR beta 5 antibody
    • MGC117330 antibody
    • MHC cell surface glycoprotein antibody
    • MHC class II antigen DRA antibody
    • MHC II antibody
    • MLRW antibody
    see all

画像

  • All lanes : Anti-HLA-DR antibody [TAL 1B5] (ab20181) at 5 µg/ml

    Lane 1 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate
    Lane 2 : Daudi whole cell lysate (ab3951)
    Lane 3 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
    Lane 4 : Human spleen tissue lysate - total protein (ab29699)
    Lane 5 : Tonsil (Human) Tissue Lysate
    Lane 6 : Human liver tissue lysate - total protein (ab29889)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 29 kDa
    Observed band size : 29,35 kDa (why is the actual band size different from the predicted?)


    Exposure time : 16 minutes
  • Ab20181 staining human normal thymus tissue. Staining is localised to cellular membranes.
    Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  • Human peripheral blood lymphocytes stained with ab20181 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (ab20181, 0.01μg/1x106 cells) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy – peripheral blood lymphocytes.

  • Anti-HLA-DR antibody [TAL 1B5] (ab20181) at 1 µg/ml + whole tissue lysate prepared from human colon at 50 µg

    Secondary
    HRP conjugated goat anti-mouse polyclonal at 1/3000 dilution
    Developed using the ECL technique

    Predicted band size : 29 kDa
    Observed band size : 33 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 seconds

    Image courtesy of an anonymous Abreview.

    See Abreview

  • IHC image of HLA-DR [TAL 1B5] staining in Human skin melanoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab20181, 0.1µg/ml, 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.

     

    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.

  • Human peripheral blood mononuclear cells isolated from whole blood were treated with BD Cytofix/Cytoperm™ kit for intracellular staining. Cells were then incubated in 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab20181, 0.005μg/1x106 cells) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150117) at 1/2000 dilution for 30 min at 4°C.

    Isotype control antibody (black line) was mouse IgG1 [15-6E10A7] (ab170190, 0.005μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of 10,000 total events were collected using a 50mW Blue laser (488nm) and 530/30 bandpass filter. Gating strategy – lymphocytes.

参考文献

This product has been referenced in:
  • Dunne MR  et al. HLA-DR expression in tumor epithelium is an independent prognostic indicator in esophageal adenocarcinoma patients. Cancer Immunol Immunother 66:841-850 (2017). IHC ; Human . Read more (PubMed: 28315927) »
  • Ai P  et al. Tumor microenvironment contributes to Epstein-Barr virus anti-nuclear antigen-1 antibody production in nasopharyngeal carcinoma. Oncol Lett 14:2458-2462 (2017). Read more (PubMed: 28781683) »

See all 16 Publications for this product

レビューと Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Dako Flex peroxidase as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: RT°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: High pH
Sample
Human Tissue sections (Malignant melanoma/lymphocytes)
Specification
Malignant melanoma/lymphocytes
Permeabilization
No
Fixative
10% buffered formalin
Username

Abcam user community

Verified customer

投稿 Jan 16 2015

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (colon)
Specification
colon
Fixative
Acetone
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 Mar 01 2013

Thank you for contacting us. Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guaran...

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Monkey Tissue sections (Japanese macaque brain)
Specification
Japanese macaque brain
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: DAKO antigen retrieval buffer , pressure cooked for 24 minutes
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Username

Abcam user community

Verified customer

投稿 Aug 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - whole (human colon lysate)
Loading amount
50 µg
Specification
human colon lysate
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 Feb 09 2012

Here is a link to the fractionation protocol as discussed: http://www.abcam.com/index.html?pageconfig=resource&rid=11473 I really think that enriching for the membrane fraction may help. I will also discuss with one of my colleagues.

I am sorry for the delay in responding to you. I tried to call you yesterday and today but was unable to reach you, therefore I would prefer to send an email to ensure a speedy resolution. It is very disappointing that 4 products are not workin...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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