Recombinant
RabMAb

Recombinant Anti-PD-L1 antibody [EPR20529] (ab213480)

Rabbit recombinant monoclonal PD-L1 antibody [EPR20529]. Validated in WB, IP, ICC/IF and tested in Mouse. Cited in 2 publication(s). Independently reviewed in 1 review(s).

Overview

  • Product name

    Anti-PD-L1 antibody [EPR20529]
    See all PD-L1 primary antibodies
  • Description

    Rabbit monoclonal [EPR20529] to PD-L1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IP, WBmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Mouse PD-L1 aa 1-250. The exact sequence is proprietary.
    Database link: Q9EP73

  • Positive control

    • WB: Mouse placenta lysate; whole cell lysate from RAW 264.7 cells treated with 100ng/ml Interferon-gamma for 48 hours. IP: Mouse placenta lysate. ICC/IF: RAW 264.7 cells treated with IFN-? (100 ng/ml) for 24 hours.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab213480 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/IF Use at an assay dependent concentration.
IP 1/30.
WB 1/1000. Detects a band of approximately 40-60 kDa (predicted molecular weight: 33 kDa).

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

  • All lanes : Anti-PD-L1 antibody [EPR20529] (ab213480) at 1/1000 dilution

    Lane 1 : Untreated RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 2 : Whole cell lysate from RAW 264.7 treated with 100ng/ml Interferon-gamma for 48 hours

    Lysates/proteins at 10 µg per lane.

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

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


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    IFN-γ treatment induces PD-L1 expression. The molecular weight observed is consistent with what has been described in the literature. PMID: 15353579.

  • Ab213480 staining PD-L1 in RAW264.7 from Mouse macrophage cell line transformed with Abelson murine leukemia virus by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% Methanol. Samples were incubated with ab213480 at 0.4 μg/ml. AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as the secondary antibody at 2 μg/ml. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)(ab196889) was used as counterstain at 2.5 μg/ml. DAPI used as nuclear counterstain. Confocal image showing membranous staining on RAW 264.7 cells treated with IFN-γ (100 ng/ml) for 24 hours.

  • PD-L1 was immunoprecipitated from 0.35 mg of Mouse placenta lysate with ab213480 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab213480 at 1/500 dilution.

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

    Lane 1: Mouse placenta lysate, 10 µg (Input).

    Lane 2: ab213480 IP in mouse placenta lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213480 in mouse placenta lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

  • Anti-PD-L1 antibody [EPR20529] (ab213480) at 1/1000 dilution + Mouse placenta lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular weight observed is consistent with what has been described in the literature. PMID: 15353579.

References

This product has been referenced in:

  • Patel MR  et al. JAK/STAT inhibition with ruxolitinib enhances oncolytic virotherapy in non-small cell lung cancer models. Cancer Gene Ther N/A:N/A (2019). Read more (PubMed: 30622322) »
  • Poggio M  et al. Suppression of Exosomal PD-L1 Induces Systemic Anti-tumor Immunity and Memory. Cell 177:414-427.e13 (2019). Read more (PubMed: 30951669) »
See all 4 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
100 ng/ml IFN-gamma
Specification
mouse hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

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Verified customer

Submitted Jun 21 2018

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