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    products/primary-antibodies/pd-l1-antibody-cal10-bsa-and-azide-free-ab251611.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] - BSA and Azide free (ab251611)

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Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Western blot - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Western blot - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Western blot - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Immunoprecipitation - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • ELISA - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
  • Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)

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 - BSA and Azide free
  • Suitable for: ELISA, IHC-P, mIHC, WB, ICC/IF, IP
  • Knockout validated
  • Reacts with: Human, Recombinant fragment

Conjugates logo Related conjugates and formulations

Alexa Fluor® 555 Alexa Fluor® 647 Unconjugated

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Overview

  • Product name

    Anti-PD-L1 antibody [CAL10] - BSA and Azide free
    See all PD-L1 primary antibodies
  • Description

    Rabbit monoclonal [CAL10] to PD-L1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ELISA, IHC-P, mIHC, WB, ICC/IF, IPmore 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. mIHC: Human tonsil tissue and Human breast cancer 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.
  • General notes

    ab251611 is the carrier-free version of ab237726.

    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.

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
  • 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] (ab237726)
    • Alexa Fluor® 647 Anti-PD-L1 antibody [CAL10] (ab267565)
    • Alexa Fluor® 555 Anti-PD-L1 antibody [CAL10] (ab307643)
  • Compatible Secondaries

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

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)
  • Recombinant Protein

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab251611 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.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
mIHC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 33 kDa.
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
Notes
ELISA
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
mIHC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 33 kDa.
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
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] - BSA and Azide free (ab251611)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)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 was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

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

    • Western blot - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Western blot - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      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


      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

    • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)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 was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

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

    • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)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 was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

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

    • Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Immunocytochemistry/ Immunofluorescence - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

    • Western blot - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Western blot - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      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


      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

      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] - BSA and Azide free (ab251611)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)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 was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

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

    • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)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 was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

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

    • Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Multiplex immunohistochemistry - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)

      This data was developed using the same antibody clone in a different buffer formulation (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] - BSA and Azide free (ab251611)
      Western blot - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      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


      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

    • Immunoprecipitation - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Immunoprecipitation - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)

      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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237726).

    • ELISA - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      ELISA - Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)

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

    • Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)
      Anti-PD-L1 antibody [CAL10] - BSA and Azide free (ab251611)

    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 (1)

    Publishing research using ab251611? Please let us know so that we can cite the reference in this datasheet.

    ab251611 has been referenced in 1 publication.

    • Sami E  et al. The Immunosuppressive Microenvironment in BRCA1-IRIS-Overexpressing TNBC Tumors Is Induced by Bidirectional Interaction with Tumor-Associated Macrophages. Cancer Res 80:1102-1117 (2020). PubMed: 31911557

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