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    products/primary-antibodies/pd-l1-antibody-epr19759-ab213524.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 [EPR19759] (ab213524)

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  • Certificate of Compliance
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Flow Cytometry (Intracellular) - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Immunoprecipitation - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
  • Anti-PD-L1 antibody [EPR19759] (ab213524)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19759] to PD-L1
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free HRP

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Human CD274 (PD-L1) knockout A549 cell lysate (ab256831)
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Human CD274 (PD-L1) knockout A549 cell lysate (ab256866)
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Product image
Human CD274 (PD-L1) knockout A549 cell line (ab267054)

View more associated products

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 (Intra), WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type A549 treated with 100 ng/mL IFN gamma (ab259377) for 48 h cell lysate; 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 (intra): CHO-PDL1.
  • 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

    EPR19759
  • 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® 488 Anti-PD-L1 antibody [EPR19759] (ab214958)
    • Alexa Fluor® 647 Anti-PD-L1 antibody [EPR19759] (ab215251)
    • HRP Anti-PD-L1 antibody [EPR19759] (ab215349)
    • Alexa Fluor® 594 Anti-PD-L1 antibody [EPR19759] (ab216742)
    • Alexa Fluor® 568 Anti-PD-L1 antibody [EPR19759] (ab220314)
    • Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
    • Anti-PD-L1 antibody [EPR19759] - Low endotoxin, Azide free (ab246695)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CD274 (PD-L1) knockout A549 cell line (ab267054)
    • Human CD274 (PD-L1) knockout A549 cell line (ab267055)
  • KO cell lysates

    • Human CD274 (PD-L1) knockout A549 cell lysate (ab256831)
    • Human CD274 (PD-L1) knockout A549 cell lysate (ab256866)
  • Positive Controls

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Recombinant Protein

    • Recombinant human PD-L1 protein (Active) (ab167713)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Universal HIER antigen retrieval reagent (10X) (ab208572)
    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
    • Anti-PD1 antibody [NAT105] (ab52587)

Applications

The Abpromise guarantee

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 (Intra)
1/500.
WB (1)
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.
Notes
Flow Cyt (Intra)
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.
  • 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: 29126 Human
    • Omim: 605402 Human
    • SwissProt: Q9NZQ7 Human
    • Unigene: 521989 Human
    • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human spleen tissue labelling PDI with ab243644 at 1.02 µg/mL (B), PD-L 1 with ab213524 at 1/100 dilution (C) and CD68 with ab213363 at 1/300 dilution (D). Anti-Rabbit and Mouse Polymer HRP was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. Heat mediated antigen retrieval (Leica ER2, PH9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibodies from the previous round, to avoid any cross-reactivity. 

      Panel A: merged staining of anti- PD1 (green, Opal™520), anti- PD-L1 (red, Opal™570) and anti- CD68 (yellow, Opal™690).

      Panel B: Anti- PD1 stained on antigen-stimulated T cells.

      Panel C: anti- PD-L1 stained on cells involved in T cell inhibition

      Panel D: anti-CD68 stained on macrophages.

      The section was incubated in three rounds of staining: in the order of ab243644, ab213363 and ab213524 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

      Lane 1 : Wild-type A549 treated with 100 ng/ml IFN gamma (ab259377) for 48 h cell lysate
      Lanes 2 & 6 : CD274 knockout A549 treated with 100 ng/ml IFN gamma (ab259377) for 48 h cell lysate
      Lane 3 : U-87 MG cell lysate
      Lane 4 : MCF7 cell lysate
      Lane 5 : Wild-type A549 untreated cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 33 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?



      Lanes 1 - 6: Merged signal (red and green). Green - ab213524 observed at 50 kDa. Red - loading control, ab8245 observed at 37 kDa.

      ab213524 Recombinant Anti-PD-L1 antibody [EPR19759] was shown to specifically react with PD-L1 in wild-type A549 treated with 100 ng/mL IFN gamma for 48 h cells in western blot. Loss of signal was observed when both treated and untreated knockout cell line ab267054 (treated and untreated knockout cell lysates ab256831) were used. Wild-type and PD-L1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab213524 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)

      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).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [EPR19759] (ab213524)

      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.

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

      Lane 1 : Wild-type A549 treated with 100 ng/mL IFN gamma for 48 h cell lysate
      Lane 2 : CD274 knockout A549 treated with 100 ng/mL IFN gamma for 48 h cell lysate
      Lane 3 : U-87 MG cell lysate
      Lane 4 : MCF7 cell lysate
      Lane 5 : Wild-type A549 untreated cell lysate
      Lane 6 : CD274 knockout A549 untreated cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 33 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?



      Lanes 1 - 6: Merged signal (red and green). Green - ab213524 observed at 50 kDa. Red - loading control, ab8245 observed at 37 kDa.

      ab213524 Recombinant Anti-PD-L1 antibody [EPR19759] was shown to specifically react with PD-L1 in wild-type A549 treated with 100 ng/mL IFN gamma for 48 h cells in western blot. Loss of signal was observed when both treated and untreated knockout cell line ab267055 (treated and untreated knockout cell lysates ab256866) were used. Wild-type and PD-L1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab213524 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

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

      Lane 1 : H1975 (Human non-small cell lung cancer epithelial cell) whole cell lysate
      Lane 2 : NCI-H1299 (Human lung carcinoma epithelial cell) whole cell lysate
      Lane 3 : A549 (Human lung carcinoma epithelial cell) whole cell lysate
      Lane 4 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate
      Lane 5 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
      Lane 6 : SK-BR-3 (Human breast adenocarcinoma epithelial cell) whole cell lysate
      Lane 7 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
      Lane 8 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate
      Lane 9 : PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate
      Lane 10 : DU 145 (Human prostate carcinoma epithelial cell) whole cell lysate
      Lane 11 : A375 (Human malignant melanoma epithelial cell) whole cell lysate
      Lane 12 : MeWo (Human malignant melanoma fibroblast) whole cell lysate
      Lane 13 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate
      Lane 14 : Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
      Lane 15 : HepG2(Human hepatocellular carcinoma epithelial cell) whole cell lysate
      Lane 16 : BXPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate
      Lane 17 : PANC-1 (Human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate
      Lane 18 : NIH:OVCAR-3 (Human ovary adenocarcinoma epithelial cell) whole cell lysate
      Lane 19 : SK-OV-3 (Human ovarian cancer epithelial cell) whole cell lysate
      Lane 20 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

      Developed using the ECL technique.

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


      Exposure time: 180 seconds


      Expression of PD-L1 varied widely among the tumor cell lines

      Blocking buffer and concentration : 5% NFDM/TBST

      Diluting buffer and concentration : 5% NFDM/TBST

    • Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
      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 (Intracellular) - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Flow Cytometry (Intracellular) - Anti-PD-L1 antibody [EPR19759] (ab213524)

      Intracellular 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. 

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [EPR19759] (ab213524)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)

      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)

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)

      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).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
      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.

    • Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Western blot - Anti-PD-L1 antibody [EPR19759] (ab213524)
      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).

    • Immunoprecipitation - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Immunoprecipitation - Anti-PD-L1 antibody [EPR19759] (ab213524)

      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 Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] (ab213524)

      Tissue Microarrays stained for "Anti-PD-L1 antibody [EPR19759]” using "ab213524" 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 perform heat mediated antigen retrieval before commencing with IHC staining protocol. The sections were incubated with ab213524 at +4°C overnight. The secondary antibody is rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Anti-PD-L1 antibody [EPR19759] (ab213524)
      Anti-PD-L1 antibody [EPR19759] (ab213524)

    Protocols

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

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

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    References (41)

    Publishing research using ab213524? Please let us know so that we can cite the reference in this datasheet.

    ab213524 has been referenced in 41 publications.

    • Xu S  et al. LSD1 silencing contributes to enhanced efficacy of anti-CD47/PD-L1 immunotherapy in cervical cancer. Cell Death Dis 12:282 (2021). PubMed: 33731702
    • 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 Y  et al. Extracellular vesicles derived from lung cancer cells exposed to intermittent hypoxia upregulate programmed death ligand 1 expression in macrophages. Sleep Breath N/A:N/A (2021). PubMed: 34254261
    • Wu SY  et al. MYC suppresses STING-dependent innate immunity by transcriptionally upregulating DNMT1 in triple-negative breast cancer. J Immunother Cancer 9:N/A (2021). PubMed: 34321275
    • Chen Z  et al. LncRNA SOX2-OT/miR-30d-5p/PDK1 Regulates PD-L1 Checkpoint Through the mTOR Signaling Pathway to Promote Non-small Cell Lung Cancer Progression and Immune Escape. Front Genet 12:674856 (2021). PubMed: 34394184
    View all Publications for this product

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    Western blot abreview for Anti-PD-L1 antibody [EPR19759]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    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.

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    Submitted Nov 12 2018

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