Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20115] to CD276 - BSA and Azide free
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-CD276 antibody [EPR20115] - BSA and Azide free
    See all CD276 primary antibodies

  • Description

    Rabbit monoclonal [EPR20115] to CD276 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment aa 1-400. The exact sequence is proprietary.
    Database link: Q5ZPR3

  • Positive control

    • WB: LNCaP, U-2OS, PC-3, 293T and MDA-MB-435S whole cell lysates; Human lung, skin and placenta lysates. IHC-P: Human tonsil, placenta, hepatocellular carcinoma, lung carcinoma and breast cancer tissues.

  • General notes

    Ab227577 is the carrier-free version of ab219648. 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.

    ab227577 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab227577 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 with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB Use at an assay dependent concentration. Detects a band of approximately 90-110 kDa (predicted molecular weight: 57 kDa).

Target

  • Function

    May participate in the regulation of T-cell-mediated immune response. May play a protective role in tumor cells by inhibiting natural-killer mediated cell lysis as well as a role of marker for detection of neuroblastoma cells. May be involved in the development of acute and chronic transplant rejection and in the regulation of lymphocytic activity at mucosal surfaces. Could also play a key role in providing the placenta and fetus with a suitable immunological environment throughout pregnancy. Both isoform 1 and isoform 2 appear to be redundant in their ability to modulate CD4 T-cell responses. Isoform 2 is shown to enhance the induction of cytotoxic T-cells and selectively stimulates interferon gamma production in the presence of T-cell receptor signaling.

  • Tissue specificity

    Ubiquitous but not detectable in peripheral blood lymphocytes or granulocytes. Weakly expressed in resting monocytes. Expressed in dendritic cells derived from monocytes. Expressed in epithelial cells of sinonasal tissue. Expressed in extravillous trophoblast cells and Hofbauer cells of the first trimester placenta and term placenta.

  • Sequence similarities

    Belongs to the immunoglobulin superfamily. BTN/MOG family.
    Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 2 Ig-like V-type (immunoglobulin-like) domains.

  • Cellular localization

    Membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • 4Ig B7 H3 antibody
      • 4Ig-B7-H3 antibody
      • AU016588 antibody
      • B7 H3 antibody
      • B7 homolog 3 antibody
      • B7-H3 antibody
      • B7H3 antibody
      • B7RP-2 antibody
      • CD_antigen=CD276 antibody
      • CD276 antibody
      • CD276 antigen antibody
      • CD276 molecule antibody
      • CD276_HUMAN antibody
      • Costimulatory molecule antibody
      • Flags: Precursor antibody
      • PSEC0249 antibody
      • UNQ309/PRO352 antibody
      see all

    Images

    • Western blot - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)
      Western blot - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)
      All lanes : Anti-CD276 antibody [EPR20115] (ab219648) at 1/1000 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : CD276 knockout HEK293T cell lysate
      Lane 3 : LNCaP cell lysate
      Lane 4 : Raji cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 57 kDa
      Observed band size: 90-110 kDa
      why is the actual band size different from the predicted?



      This data was developed using the same antibody clone in a different buffer formulation (ab219648).

      Lanes 1-4: Merged signal (red and green). Green - ab219648 observed at 90-110 kDa. Red - loading control ab8245 observed at 36 kDa.

      ab219648 Anti-CD276 antibody [EPR20115] was shown to specifically react with CD276 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266658 (knockout cell lysate ab257097) was used. Wild-type and CD276 knockout samples were subjected to SDS-PAGE. ab219648 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

      Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Membranous staining on human hepatocellular carcinoma is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219648).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

      Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Membranous staining on human lung carcinoma is observed [PMID: 25609202].

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219648).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

      Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Membranous and cytoplasmic staining on human breast cancer tumor cells and stromal cells is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219648).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

      This IHC data was generated using the same anti-CD276 antibody clone, EPR20115, in a different buffer formulation (cat# ab219648).

      Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Membranous staining on dendritic cells of human tonsil is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

      This IHC data was generated using the same anti-CD276 antibody clone, EPR20115, in a different buffer formulation (cat# ab219648).

      Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Cytoplasmic and membranous staining on vascular endothelial cells and trophoblast cells of human placenta is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)

      Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Negative tissue: no staining on human skeletal muscle. [PMID: 18975328].

      Counter stained with Hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219648).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    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

    References (0)

    Publishing research using ab227577? Please let us know so that we can cite the reference in this datasheet.

    ab227577 has not yet been referenced specifically in any publications.

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