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    products/primary-antibodies/pd-l1-antibody-cal10-ab237726.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 [CAL10] (ab237726)

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Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Immunoprecipitation - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
  • ELISA - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
  • Anti-PD-L1 antibody [CAL10] (ab237726)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [CAL10] to PD-L1
  • Suitable for: ELISA, WB, ICC/IF, IP, mIHC, IHC-P
  • Knockout validated
  • Reacts with: Human, Recombinant fragment

Conjugates logo Related conjugates and formulations

Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free

You may also be interested in

Protein
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Recombinant human PD-L1 protein (Active) (ab167713)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Primary
Product image
Anti-PD-L1 antibody [EPR19759] - BSA and Azide free (ab221612)

View more associated products

Overview

  • Product name

    Anti-PD-L1 antibody [CAL10]
    See all PD-L1 primary antibodies
  • Description

    Rabbit monoclonal [CAL10] to PD-L1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ELISA, WB, ICC/IF, IP, mIHC, IHC-Pmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Human, Recombinant fragment
  • Immunogen

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

  • Positive control

    • IHC-P: Human NSCLC tissue; PD L1 transfected HEK-293 cells. Human lung carcinoma and placenta tissue. WB: CHO-PD-L1 (PD-L1 stably expressed Chinese hamster ovary epithelial cell) whole cell lysate. A549 treated with 100ng/ml IFN gamma for 48h whole cell lysate. Human placenta lysate. ICC/IF: CHO-PD-L1 (PD-L1 stably expressed Chinese hamster ovary epithelial cell) cells. IP: ab237726 IP in NCI-H1975 whole cell lysate. mIHC: Human tonsil tissue and Human breast cancer tissue.

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: 0.05% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Purification notes

    Purity >99%
  • Clonality

    Monoclonal
  • Clone number

    CAL10
  • 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 [CAL10] - BSA and Azide free (ab251611)
    • Alexa Fluor® 647 Anti-PD-L1 antibody [CAL10] (ab267565)
  • Compatible Secondaries

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

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

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

    • PD-L1 / PD1 Multiplex IHC-IF Antibody Panel (PD-L1, PD1, CD68, CD3, Ki67, panCK) (ab269812)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab237726 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
ELISA
Use at an assay dependent concentration.
WB
1/1000. Predicted molecular weight: 33 kDa.
ICC/IF
1/1000.
IP
1/30.
mIHC
Use at an assay dependent concentration.
IHC-P
1/1000 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
ELISA
Use at an assay dependent concentration.
WB
1/1000. Predicted molecular weight: 33 kDa.
ICC/IF
1/1000.
IP
1/30.
mIHC
Use at an assay dependent concentration.
IHC-P
1/1000 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Application notes
Is unsuitable for Flow Cyt.

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

    • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
      Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
      All lanes : Anti-PD-L1 antibody [CAL10] - Mouse IgG1 (ab279292) at 1/1000 dilution

      Lane 1 : Wild-type A549 Treated IFN-gamma (100 ng/ml) for 48 hours cell lysate
      Lane 2 : CD274 knockout A549 Treated IFN-gamma (100 ng/ml) for 48 hours cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

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



      False colour image of Western blot: Anti-PD-L1 antibody [CAL10] - Mouse IgG1 staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab279292 was shown to bind specifically to PD-L1. A band was observed at 48 kDa in treated wild-type A549 cell lysates with no signal observed at this size in Cd274 knockout cell line ab267054 (knockout cell lysate ab256831). To generate this image, wild-type and Cd274 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

    • Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
      Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
      All lanes : Anti-PD-L1 antibody [CAL10] (ab237726) at 1/1000 dilution

      Lane 1 : CHO-S (Chinese hamster ovary epithelial cell) whole cell lysate
      Lane 2 : CHO-PD-L1 (PD-L1 stably expressed Chinese hamster ovary epithelial cell) 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


      Exposure time: 3 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)

      IHC image of ab237726 staining PD-L1 in PD-L1 Dynamic Range Analyte Control formalin fixed paraffin embedded cell lines (HistoCyte Laboratories), performed on a Leica BOND RX (standard Protocol F, Polymer Refine kit). The section was pre-treated using heat mediated antigen retrieval with sodium  citrate buffer (pH6, epitope retrieval solution 1) for 30 mins at 98°C. The section was then incubated with ab237726, 1μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system for 8 minutes at room temperature. DAB was used  as the chromogen for 10 minutes at room temperature. The section was then counterstained with hematoxylin, blued, dehydrated, cleared and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

       

    • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [CAL10] (ab237726)
      Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [CAL10] (ab237726)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized CHO-PD-L1 (PD-L1 stably expressed Chinese hamster ovary epithelial cell) and CHO-S (chinese hamster ovary epithelial cell) cells labeling PD-L1 with ab237726 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining in CHO-PD-L1 cells. The nuclear counter stain is DAPI (blue). Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red). The negative control is the secondary antibody only.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)

      IHC image of ab237726 staining PD-L1 in human tonsil formalin fixed paraffin embedded tissue sections*, performed on a Leica BOND RX (standard Protocol F, Polymer Refine kit). The section was pre-treated using heat mediated antigen retrieval with sodium  citrate buffer (pH6, epitope retrieval solution 1) for 30 mins at 98°C. The section was then incubated with ab237726, 1μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system for 8 minutes at room temperature. DAB was used  as the chromogen for 10 minutes at room temperature. The section was then counterstained with hematoxylin, blued, dehydrated, cleared and mounted with DPX.

       

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

       

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
      Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
      All lanes : Anti-PD-L1 antibody [CAL10] (ab237726) at 1/1000 dilution

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


      Exposure time: 26 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The molecular mass observed is consistent with what has been described in the literature (PMID: 26546452).

    • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] (ab237726)

      Fluorescence multiplex immunohistochemical analysis of normal human tonsil tissue (formalin-fixed paraffin-embedded section).

      Merged staining of anti-PD1 (ab237728; orange; Opal™520), anti-PDL1 (ab237726; green; Opal™540), anti-CD68 (ab192847; yellow; Opal™570), anti-CD3 (ab16669; red; Opal™620), anti-Ki67 (ab16667; light blue; Opal™650) and anti-PanCK (ab7753; grey; Opal™690).

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 7-color automation IHC kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; in the order of ab237728 (1/500 dilution), ab237726 (1/500 dilution), ab192847 (1/300 dilution), ab16669 (1/300 dilution), ab16667 (1/200 dilution) and ab7753 (1/200 dilution); each using a separate fluorescent tyramide signal amplification system.

      Sodium citrate antigen retrieval (Leica ER1, pH6.0, 30 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

      DAPI (dark blue) was used as a nuclear counter stain.

      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra Polaris.

    • Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
      Western blot - Anti-PD-L1 antibody [CAL10] (ab237726)
      Anti-PD-L1 antibody [CAL10] (ab237726) at 1/1000 dilution + Human placenta lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 33 kDa


      Exposure time: 26 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)

      Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling PD-L1 with ab237726 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Mainly membranous staining on the human lung carcinoma. The section was incubated with ab237726 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 30 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)

      Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling PD-L1 with ab237726 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on the human placenta. The section was incubated with ab237726 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 30 mins.

    • Immunoprecipitation - Anti-PD-L1 antibody [CAL10] (ab237726)
      Immunoprecipitation - Anti-PD-L1 antibody [CAL10] (ab237726)

      PD-L1 was immunoprecipitated from 0.35 mg NCI-H1975 (human non-small cell lung cancer cell line) whole cell lysate with ab237726 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237726 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.
      Lane 1: NCI-H1975 whole cell lysate 10 µg (Input).
      Lane 2: ab237726 IP in NCI-H1975 whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237726 in NCI-H1975 whole cell lysate.
      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 30 seconds.

      The molecular mass observed is consistent with what has been described in the literature (PMID: 26546452).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)

      Formalin-fixed, paraffin-embedded NSCLC (Non-small-cell lung carcinoma) tissue stained for PD L1 using ab237726 at 0.3 µg/ml dilution in immunohistochemical analysis. Positive staining (Left panel) and negative staining (Right panel).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)

      Formalin-fixed, paraffin-embedded PD L1 transfected HEK-293 (Human epithelial cell line from embryonic kidney) cells stained for PD L1 using ab237726 at 0.3 µg/ml dilution (Left panel) in immunohistochemical analysis.

      Negative control (Right panel): PD L2 transfected HEK-293 cells.

    • ELISA - Anti-PD-L1 antibody [CAL10] (ab237726)
      ELISA - Anti-PD-L1 antibody [CAL10] (ab237726)

      ELISA - Anti-PD-L1 antibody [CAL10] (ab237726).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] (ab237726)

      Tissue Microarrays stained for "Anti-PD-L1 antibody [CAL10]” using "ab237726" 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 pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 30 minutes. The sections were incubated with ab237726 for 15 mins at room temperature followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Anti-PD-L1 antibody [CAL10] (ab237726)
      Anti-PD-L1 antibody [CAL10] (ab237726)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (9)

    Publishing research using ab237726? Please let us know so that we can cite the reference in this datasheet.

    ab237726 has been referenced in 9 publications.

    • Mei J  et al. A comparability study of natural and deglycosylated PD-L1 levels in lung cancer: evidence from immunohistochemical analysis. Mol Cancer 20:11 (2021). IHC-P ; Human . PubMed: 33413365
    • Yao X  et al. Endoplasmic reticulum stress-induced exosomal miR-27a-3p promotes immune escape in breast cancer via regulating PD-L1 expression in macrophages. J Cell Mol Med 24:9560-9573 (2020). PubMed: 32672418
    • Zhou S  et al. Evaluation of PD-1 blockade using a multicellular tumor spheroid model. Am J Transl Res 11:7471-7478 (2019). PubMed: 31934294
    • Xue D  et al. IL-21/IL-21R inhibit tumor growth and invasion in non-small cell lung cancer cells via suppressing Wnt/ß-catenin signaling and PD-L1 expression. Int J Mol Med 44:1697-1706 (2019). PubMed: 31573051
    • Pichler R  et al. Prognostic Value of Testing PD-L1 Expression After Radical Cystectomy in High-risk Patients. Clin Genitourin Cancer N/A:N/A (2018). IHC-P ; Human . PubMed: 29960831
    • Pratt D  et al. Programmed Death Ligand 1 Is a Negative Prognostic Marker in Recurrent Isocitrate Dehydrogenase-Wildtype Glioblastoma. Neurosurgery N/A:N/A (2018). IHC-P ; Human . PubMed: 30011045
    • Karnik T  et al. PD-L1 in breast cancer: comparative analysis of 3 different antibodies. Hum Pathol 72:28-34 (2018). IHC-P ; Human . PubMed: 28843709
    • Zarogoulidis P  et al. Immunotherapy "Shock" a case series of PD-L1 100% and pembrolizumab first-line treatment. Respir Med Case Rep 22:197-202 (2017). IHC-P ; Human . PubMed: 28879076
    • Hutarew G PD-L1 testing, fit for routine evaluation? From a patholo?gist's point of view. Memo 9:201-206 (2016). PubMed: 28058063

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