Recombinant
RabMAb

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

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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: IHC-P, IP, ICC/IF, WB, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human PD-L1 aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9NZQ7

  • Positive control

    • WB: 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. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab221612 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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

Applications

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
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.
ICC/IF 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 polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) (ab150077).
Flow Cyt 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.
  • 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

    • 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)

      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.

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

      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).

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

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

    • 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).

    • 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.

    References

    ab221612 has not yet been referenced specifically in any publications.

    Publishing research using ab221612? Please let us know so that we can cite the reference in this datasheet.

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