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    pd-l1-antibody-epr19759-bsa-and-azide-free-ab221612.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] - BSA and Azide free (ab221612)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Western blot - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Flow Cytometry (Intracellular) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Western blot - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Immunoprecipitation - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
  • Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

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 - BSA and Azide free
  • Suitable for: ICC, IHC-P, IP, WB, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 HRP Unconjugated

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Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
Primary
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Anti-PD-L1 antibody [EPR19759] (ab213524)

View more associated products

Overview

  • Product name

    Anti-PD-L1 antibody [EPR19759] - BSA and Azide free
    See all PD-L1 primary antibodies
  • Description

    Rabbit monoclonal [EPR19759] to PD-L1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC, IHC-P, IP, WB, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab150077)

  • 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.
  • General notes

    ab221612 is the carrier-free version of ab213524.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • 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

    • Anti-PD-L1 antibody [EPR19759] (ab213524)
    • 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] - Low endotoxin, Azide free (ab246695)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Immunizing Peptide (Blocking)

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

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

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Universal HIER antigen retrieval reagent (10X) (ab208572)
    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab221612 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
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

 

IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 40-45 kDa (predicted molecular weight: 33 kDa).Can be blocked with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077).
Flow Cyt (Intra)
Use at an assay dependent concentration.
Notes
ICC
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

 

IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 40-45 kDa (predicted molecular weight: 33 kDa).Can be blocked with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077).
Flow Cyt (Intra)
Use at an assay dependent concentration.

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] - BSA and Azide free (ab221612)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

      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] - BSA and Azide free (ab221612)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

      This IHC data was generated using the same anti-PDL1 antibody clone, EPR19759, in a different buffer formulation (cat# 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).

      Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    • Western blot - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      Western blot - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      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
      Lane 2 : 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
      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?



      This data was developed using the same antibody clone in a different buffer formulation (ab213524).

      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.

    • Flow Cytometry (Intracellular) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      Flow Cytometry (Intracellular) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

      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. 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213524). 

       

       

    • Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

      Clone EPR19759 (ab221612) has been successfully conjugated by Abcam. This image was generated using Anti-PD-L1 antibody [EPR19759] (Alexa Fluor® 647). Please refer to ab215251 for protocol details.

      ab215251 staining PDL1 in CHO-PDL1 cells. The lower panels demonstrate that ab215251 does not cross react with un-transfected CHO cells.

      The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab215251 at 1/200 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

      Clone EPR19759 (ab221612) has been successfully conjugated by Abcam. This image was generated using Anti-PD-L1 antibody [EPR19759] (Alexa Fluor® 488). Please refer to ab214958 for protocol details.

      ab214958 staining PDL1 in CHO-PDL1 cells. The lower panels demonstrate that ab214958 does not cross react with un-transfected CHO cells.

      The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab214958 at 1/200 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

      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)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213524).

      Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    • Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213524).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

      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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213524).

      Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    • Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      Immunocytochemistry - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213524).

    • Western blot - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      Western blot - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      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?



      This data was developed using the same antibody clone in a different buffer formulation (ab213524).

      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.

    • Immunoprecipitation - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      Immunoprecipitation - Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

      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 1/10000 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213524).

    • Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)
      Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

    Protocols

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    References (0)

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    ab221612 has not yet been referenced specifically in any publications.

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