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, IHC-Pmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is considered to be commercially sensitive.

  • 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.

Properties

Applications

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.
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.
    • Information by UniProt
    • Database links
    • Alternative names
      • B7 H antibody
      • B7 H1 antibody
      • B7 homolog 1 antibody
      • B7-H1 antibody
      • B7H antibody
      • B7H1 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

    • 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

    • 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.

       

    • 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).

    • 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.

    • 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.

    • 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.

    • 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.

    • 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.

    • 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).

    • 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).

    • ELISA - 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.

    References

    This product has been referenced in:
    • 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 . Read more (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 . Read more (PubMed: 30011045) »
    See all 5 Publications for this product

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