Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

Overview

  • Product nameAnti-PD-L1 antibody [28-8] - Low endotoxin, Azide free
    See all PD-L1 primary antibodies
  • Description
    Rabbit monoclonal [28-8] to PD-L1 - Low endotoxin, Azide free
  • Tested applicationsSuitable for: IHC-P, Flow Cyt, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein within Human PD-L1. The exact sequence is proprietary.
    Database link: Q9NZQ7

  • Positive control
    • Tissue: Human tonsil and head and neck squamous cell carcinoma tissues; L2987 cell line. Cell Lines: Positives: B-CPAP- high, ES-2- medium, HCC70 - low For additional information, please refer to here: Programmed death-ligand 1 (PD-L1) expression in various tumor types - http://www.immunotherapyofcancer.org/content/1/S1/P53
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Anti-PD-L1 antibody [28-8] has been used as detector antibody in Human PD-L1 SimpleStep ELISA® kit (ab214565)

    Additional information on positive controls:

    Tissue:
    Tonsil- with hyperreactive changes
    Note: Tonsil Specimens- is recommended to screen several hyper-reactive tonsils to find those with 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
    Note: Tumor Specimens- PD-L1 expression varies by tumor type so screening is recommended to find positive and negative tumor controls. Refer to web link publication below to find some suggested tumor types. Many tumor specimens have some inflammatory macrophages and mononuclear leukocytes. Best to look for specimens with high numbers of these cells

    Cell Lines:
    Positives: B-CPAP- high, ES-2- medium, HCC70 - low

    For primary negative control, isotype control, RabMAb negative control antibody (Ab172730) is recommended.

    For negative control sample, cell line COLO205 is recommended.

    For PD-L1 protein, see ab167713

    Recommended protocols:

    For recommended Western Blotting (WB) protocol, please refer to the protocol book (line 3) in the protocol section.

    For recommended Immunohistochemistry (IHC) protocol, please refer to the protocol book (line 2) in the protocol section and/or here (downloadable copy)

    For IHC usage on FFPE tissues, the following antigen solution is recommended with clone 28-8 -  Universal HIER antigen retrieval reagent (ab208572) KO Validated

    For recommended Flow Cytometry (Flow Cyt) protocol, please refer to the protocol book (line 1) in the protocol section and/or here (downloadable copy)

    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. 
    For colormetric detection of PDL1, it is recommended to use Anti-PD-L1 antibody (ab58810)

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

Properties

Applications

Our Abpromise guarantee covers the use of ab209889 in the following tested applications.

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

Application Abreviews Notes
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB Use at an assay dependent concentration. Detects a band of approximately 33-43 kDa (predicted molecular weight: 33 kDa).

Please check the parent abID, ab205921, for more information on dilutions.

Target

  • FunctionInvolved 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.
  • Tissue specificityHighly 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.
  • Sequence similaritiesBelongs 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.
  • Cellular localizationCell membrane and Endomembrane system.
  • Information by UniProt
  • Database links
  • Alternative names
    • B7 H antibody
    • B7 H1 antibody
    • B7 homolog 1 antibody
    • B7-H1 antibody
    • B7H antibody
    • B7H1 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

Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free images

  • Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889) + NCI-H1975 (human non-small cell lung cancer) whole cell lysate at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

    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 buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM/TBST

  • All lanes : Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

    Lane 1 : CHO-S cell lysate
    Lane 2 : Human PD-L1 transfected CHO-S cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

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


    Exposure time : 1 minute

    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM/TBST

     

References for Anti-PD-L1 antibody [28-8] - Low endotoxin, Azide free (ab209889)

ab209889 has not yet been referenced specifically in any publications.

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