Overview

  • Product name
    Anti-PD-L1 antibody [EPR19759]
    See all PD-L1 primary antibodies
  • Description
    Rabbit monoclonal [EPR19759] to PD-L1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human PD-L1 aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9NZQ7

  • Positive control
    • WB: Chinese hamster ovary cell lysate overexpressing PD-L1; NCI-H1975 whole cell lysate. IHC-P: Human tonsil, placenta and stomach cancer tissues. ICC/IF: CHO-PDL1 and NCI-H1975 cells. IP: NCI-H1975 whole cell lysate. Flow Cyt: CHO-PDL1.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab213524 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
Flow Cyt 1/500.
WB 1/1000. Detects a band of approximately 40-60 kDa (predicted molecular weight: 33 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Antigen retrieval: Universal HIER antigen retrieval reagent (ab208572).

ICC/IF 1/500.
IP 1/30.

Target

  • Function
    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.
  • Tissue specificity
    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.
  • Sequence similarities
    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.
  • Cellular localization
    Cell 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

Images

  • Anti-PD-L1 antibody [EPR19759] (ab213524)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling PD-L1 with ab213524 at a dilution of 1:250. 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.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling PD-L1 with ab213524 at 1/250 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Membrane staining on the human tonsil crypt epithelium is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized CHO (Chinese hamster ovary cell line) cells labeling PD-L1 with ab213524 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing membrane and cytoplasmic staining on CHO-PDL1 cells.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab213524 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.

    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • All lanes : Anti-PD-L1 antibody [EPR19759] (ab213524) at 0.5 µg/ml

    Lane 1 : Untreated A549 (Human lung carcinoma epithelial cell) whole cell lysate
    Lane 2 : A549 (Human lung carcinoma epithelial cell) treated with 100ng/ml Interferon gamma for 48 hours whole cell lysate

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

    Predicted band size: 33 kDa


    Exposure time: 3 seconds


    Blocking and diluting buffer: 5% NFDM/TBST.

  • Flow Cytometry analysis of CHO-PD-L1 (red) and CHO-S (blue) cells labelling PD-L1 with ab213524 at 1/500. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% tween-20-PBS and blocked with 10% goat serum. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NCI-H1975 (Human lung non small cell carcinoma cell line) cells labeling PD-L1 with ab213524 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing weakly membrane and cytoplasmic staining on NCI-H1975 cells.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab213524 at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.

    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling PD-L1 with ab213524 at 1/250 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Membrane staining on the human placenta is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is rabbit specific IHC polymer detection kit HRP/DAB (ab209101)

  • Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue labeling PD-L1 with ab213524 at 1/250 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Membrane staining on the human stomach cancer is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • All lanes : Anti-PD-L1 antibody [EPR19759] (ab213524) at 1/1000 dilution

    Lane 1 : Chinese hamster ovary cell lysate
    Lane 2 : Chinese hamster ovary cell lysate overexpressing PD-L1

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


    Exposure time: 3 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The lower band is predicted to be isoform 2.

  • Anti-PD-L1 antibody [EPR19759] (ab213524) at 1/1000 dilution + NCI-H1975 (Human non-small cell lung cancer cell line) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 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/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 26546452).

  • PD-L1 was immunoprecipitated from 0.35 mg of NCI-H1975 (Human non-small cell lung cancer cell line) whole cell lysate with ab213524 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab213524 at 1/1000 dilution.

    VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.

    Lane 1: NCI-H1975 whole cell lysate 10µg (Input).

    Lane 2: ab213524 IP in NCI-H1975 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213524 in NCI-H1975 whole cell lysate.

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

    Exposure time: 3 minutes.

References

This product has been referenced in:
  • Chen Y  et al. Antitumor effects of the silencing of programmed cell death ligand 1 in colorectal cancer via immunoregulation. Oncol Rep 40:3370-3380 (2018). Read more (PubMed: 30272332) »
  • Liu Y  et al. PD-1 blockade inhibits lymphocyte apoptosis and restores proliferation and anti-viral immune functions of lymphocyte after CP and NCP BVDV infection in vitro. Vet Microbiol 226:74-80 (2018). Read more (PubMed: 30389046) »
See all 5 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (A549)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Treatment
IFN-gamma, 100 ng/ml, 48 hr
Specification
A549
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C

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Submitted Nov 12 2018

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

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