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    products/primary-antibodies/pd-l1-antibody-epr20529-ab213480.pdf

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Immunology Cell Type Markers CD Non-lineage
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PD-L1 antibody [EPR20529] (ab213480)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (5) Submit a question References (26)

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Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480)
  • Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480)
  • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [EPR20529] (ab213480)
  • Immunoprecipitation - Anti-PD-L1 antibody [EPR20529] (ab213480)
  • Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480)
  • Anti-PD-L1 antibody [EPR20529] (ab213480)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20529] to PD-L1
  • Suitable for: ICC/IF, IP, WB
  • Knockout validated
  • Reacts with: Mouse

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Alexa Fluor® 647 Carrier Free

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Overview

  • Product name

    Anti-PD-L1 antibody [EPR20529]
    See all PD-L1 primary antibodies
  • Description

    Rabbit monoclonal [EPR20529] to PD-L1
  • Host species

    Rabbit
  • Specificity

    ab213480 is not suitable in IHC-FR for mouse samples

  • Tested applications

    Suitable for: ICC/IF, IP, WBmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

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

  • Positive control

    • WB: Mouse placenta lysate, HEK mPDL1 cell lysate, whole cell lysate from RAW 264.7 cells treated with Interferon-gamma and whole cell lysate from MEF cells treated with Interferon-gamma. IP: Mouse placenta lysate. ICC/IF: RAW 264.7 cells treated with IFN-? (100 ng/ml) for 24 hours.
  • General notes

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20529
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Cell Type Markers
    • CD
    • Non-lineage
    • Immunology
    • Immune System Diseases
    • Autoimmune
    • Cancer
    • Tumor immunology
    • Tumor-associated antigens
    • Cancer
    • Tumor biomarkers
    • Tumor antigens

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-PD-L1 antibody [EPR20529] (ab224030)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Mouse PD-L1 protein (ab130039)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Anti-PD1 antibody [NAT105] (ab52587)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab213480 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
ICC/IF
Use at an assay dependent concentration.
IP
1/30.
WB (3)
1/1000. Detects a band of approximately 40-60 kDa (predicted molecular weight: 33 kDa).
Notes
ICC/IF
Use at an assay dependent concentration.
IP
1/30.
WB
1/1000. Detects a band of approximately 40-60 kDa (predicted molecular weight: 33 kDa).
Application notes
Is unsuitable for IHC-P.

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.
  • Target information above from: UniProt accession Q9NZQ7 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 60533 Mouse
    • SwissProt: Q9EP73 Mouse
    • Unigene: 245363 Mouse
    • Alternative names

      • B7 H antibody
      • B7 H1 antibody
      • B7 homolog 1 antibody
      • B7-H1 antibody
      • B7H antibody
      • B7H1 antibody
      • CD 274 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

    • Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480)
      Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480)
      All lanes : Anti-PD-L1 antibody [EPR20529] (ab213480) at 1/5000 dilution

      Lane 1 : Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
      Lane 2 : RAW264.7 treated with 100 ng/ml IFN gamma for 48 h, 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: 33 kDa
      Observed band size: 40-60 kDa why is the actual band size different from the predicted?


      Exposure time: 37 seconds


      This WB was performed by using YCA-R20529(BF)-111A H3L2 YR120921PJ, 1:5000 dilution. Working concentration: 0.402 μg/ml.

      Blocking buffer and concentration: 5% NFDM/TBST

      Diluting buffer and concentration: 5% NFDM/TBST

       

    • Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480)
      Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480)This image is courtesy of an anonymous collaborator.
      All lanes : Anti-PD-L1 antibody [EPR20529] (ab213480) at 100 µg

      Lane 1 : HEK mPDL1 cell lysate at 20 µg
      Lane 2 : HEK hPDL1 cell lysate at 20 µg
      Lane 3 : HEK CL7 at 20 µg
      Lane 4 : RAW 264.7 Wild-type (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 100 µg
      Lane 5 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate treated with Interferon-gamma at 100 µg
      Lane 6 : MEF Wild-type (Mouse embryonic fibroblast cell line) whole cell lysate at 100 µg
      Lane 7 : MEF (Mouse embryonic fibroblast cell line) whole cell lysate treated with Interferon-gamma at 100 µg
      Lane 8 : MEF mPDL1 KO cell lysate at 100 µg
      Lane 9 : MEF mPDL1 KO treated with Interferon-gamma at 100 µg

      Predicted band size: 33 kDa
      Observed band size: 48 kDa why is the actual band size different from the predicted?


      Exposure time: 5 minutes


      The mAb is specific for mouse mPDL1 and does not recognise hPDL1. As expected a specific 48kDa band is observed in RAW264.7 and MEFS cell extract after Interferon-gamma treatment. No bands are observed in RAW264.7 and MEFS WT and in MEFS KO with and without Interferon-gamma treatment.

      See Abreview

    • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [EPR20529] (ab213480)
      Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [EPR20529] (ab213480)

      Ab213480 staining PD-L1 in RAW264.7 from Mouse macrophage cell line transformed with Abelson murine leukemia virus by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% Methanol. Samples were incubated with ab213480 at 0.4 μg/ml. AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as the secondary antibody at 2 μg/ml. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)(ab196889) was used as counterstain at 2.5 μg/ml. DAPI used as nuclear counterstain. Confocal image showing membranous staining on RAW 264.7 cells treated with IFN-γ (100 ng/ml) for 24 hours.

    • Immunoprecipitation - Anti-PD-L1 antibody [EPR20529] (ab213480)
      Immunoprecipitation - Anti-PD-L1 antibody [EPR20529] (ab213480)

      PD-L1 was immunoprecipitated from 0.35 mg of Mouse placenta lysate with ab213480 at 1/30 dilution.

      Western blot was performed from the immunoprecipitate using ab213480 at 1/500 dilution.

      VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution.

      Lane 1: Mouse placenta lysate, 10 µg (Input).

      Lane 2: ab213480 IP in mouse placenta lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213480 in mouse placenta lysate.

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

      Exposure time: 3 seconds.

    • Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480)
      Western blot - Anti-PD-L1 antibody [EPR20529] (ab213480)
      Anti-PD-L1 antibody [EPR20529] (ab213480) at 1/1000 dilution + Mouse placenta lysate at 10 µg

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

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


      Exposure time: 15 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The molecular weight observed is consistent with what has been described in the literature. PMID: 15353579.

    • Anti-PD-L1 antibody [EPR20529] (ab213480)
      Anti-PD-L1 antibody [EPR20529] (ab213480)

    Protocols

    • Western blot protocols
    • Immunoprecipitation protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (26)

    Publishing research using ab213480? Please let us know so that we can cite the reference in this datasheet.

    ab213480 has been referenced in 26 publications.

    • Fang W  et al. Progranulin induces immune escape in breast cancer via up-regulating PD-L1 expression on tumor-associated macrophages (TAMs) and promoting CD8+ T cell exclusion. J Exp Clin Cancer Res 40:4 (2021). PubMed: 33390170
    • Li H  et al. The Beneficial Role of Sunitinib in Tumor Immune Surveillance by Regulating Tumor PD-L1. Adv Sci (Weinh) 8:2001596 (2021). PubMed: 33510997
    • Liu X  et al. Combination Chemo-Immunotherapy for Pancreatic Cancer Using the Immunogenic Effects of an Irinotecan Silicasome Nanocarrier Plus Anti-PD-1. Adv Sci (Weinh) 8:2002147 (2021). PubMed: 33747719
    • Wang Z  et al. Abrogation of USP7 is an alternative strategy to downregulate PD-L1 and sensitize gastric cancer cells to T cells killing. Acta Pharm Sin B 11:694-707 (2021). PubMed: 33777676
    • Yang J  et al. Circular RNA CHST15 Sponges miR-155-5p and miR-194-5p to Promote the Immune Escape of Lung Cancer Cells Mediated by PD-L1. Front Oncol 11:595609 (2021). PubMed: 33777742
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-5 of 5 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PD-L1 antibody [EPR20529]

    Excellent
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate
    Permeabilization
    No
    Specification
    Liver
    Blocking step
    CAS-Block as blocking agent for 30 minute(s) · Concentration: 100µg/mL · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MISS. Adi Yehezkel

    Verified customer

    Submitted Mar 04 2022

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PD-L1 antibody [EPR20529]

    Excellent
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Lymph node)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH high
    Permeabilization
    No
    Specification
    Lymph node
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Apr 25 2021

    Western blot abreview for Anti-PD-L1 antibody [EPR20529]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (machrophages and MEFs)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    100 µg
    Specification
    machrophages and MEFs
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 37°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Apr 19 2021

    Western blot abreview for Anti-PD-L1 antibody [EPR20529]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Mouse Osteosarcoma Cells)
    Gel Running Conditions
    Reduced Denaturing (5-15)
    Loading amount
    25 µg
    Specification
    Mouse Osteosarcoma Cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jun 30 2020

    Western blot abreview for Anti-PD-L1 antibody [EPR20529]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (mouse hepatocytes)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Treatment
    100 ng/ml IFN-gamma
    Specification
    mouse hepatocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jun 21 2018

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