Recombinant
RabMAb

Recombinant Anti-CD276 antibody [EPNCIR122] (ab134161)

Rabbit recombinant monoclonal CD276 antibody [EPNCIR122]. Validated in WB, IP, IHC, Flow Cyt and tested in Mouse, Human. Cited in 6 publication(s).

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Overview

  • Product name

    Anti-CD276 antibody [EPNCIR122]
    See all CD276 primary antibodies

  • Description

    Rabbit monoclonal [EPNCIR122] to CD276

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-Fr, WB, IP, Flow Cytmore details
    Unsuitable for: ICC/IF or IHC-P

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Recombinant full length protein corresponding to Human CD276. Alternate injections with Human recombinant protein and Human CD276 over-expressing CHO cells.
    Database link: Q5ZPR3
    (Peptide available as ab182198)

  • Positive control

    • 293T, LnCaP, and JAR cell lysates, CHO-K1 cells lysates transfected with human and mouse CD276.

  • General notes

    This antibody was developed as part of a collaboration between Epitomics, the National Cancer Institute's Center for Cancer Research and the lab of Brad St. Croix. View antibodies from our collaboration with the NCI Center for Cancer Research.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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 ab134161 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-Fr Use at an assay dependent concentration. PubMed: 28399408
WB 1/200. Predicted molecular weight: 57 kDa.
IP 1/10 - 1/100.
Flow Cyt 1/1000 - 1/10000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
  • Application notes
    Is unsuitable for ICC/IF or IHC-P.

    • 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 [EPNCIR122] (ab134161)
        Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Anti-CD276 antibody [EPNCIR122] (ab134161) at 1/200 dilution + CHO (hamster ovary epithelial cell) whole cell lysates transfected with human CD276 at 15 µg

        Secondary
        Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

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


        Exposure time: 3 minutes


        Blocking/dilution buffer: 5% NFDM/TBST

        Performed using purified antibody.

      • Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)

        Overlay histogram showing THP-1 (human monocytic leukemia monocyte) cells stained with ab134161 (red line). The cells were fixed with 4% paraformaldehyde and then permeabilized with 90% methanol. The cells were incubated with the antibody (ab134161) at 1/80 dilution. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution. Isotype control antibody (black line) was rabbit monoclonal IgG (ab172730) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

        Performed using purified antibody.

      • Immunoprecipitation - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Immunoprecipitation - Anti-CD276 antibody [EPNCIR122] (ab134161)

        CD276 was immunoprecipitated from 10 ug of HEK 293 (human embryonic kidney epithelial cell) whole cell lysate with ab134161 at 1/40 diltuion. Western blot was permformed from the immunoprecipitate using ab 134161 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/500 dilution.

        Lane 1: HEK 293 whole cell lysate 10 ug (Input).

        Lane 2: ab134161 IP in HEK 293 whole cell lysate.

        Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab134161 in HEK 293 whole cell lysate.

        Blocking/dilution buffer: 5% NFDM/TBST

        Performed using purified antibody.

      • Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Anti-CD276 antibody [EPNCIR122] (ab134161) at 1/1000 dilution + LNCap (human prostate carcinoma epithelial cell) whole cell lysate at 15 µg

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

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


        Exposure time: 3 minutes


        Blocking/dilution buffer: 5% NFDM/TBST

        Performed using purified antibody.

      • Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)

        Overlay histogram showing THP1 cells stained with ab134161 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% human serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab134161, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

      • Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
        All lanes : Anti-CD276 antibody [EPNCIR122] (ab134161) at 0.264 µg/ml

        Lane 1 : CHO-K1 cell lysates, transfected with Human CD276
        Lane 2 : CHO-K1 cell lysates, transfected with Mouse CD276.

        Lysates/proteins at 20 µg per lane.

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

        Predicted band size: 57 kDa


        Exposure time: 3 minutes


        Blocking/dilution buffer: 5% NFDM/TBST

      • Immunohistochemistry (Frozen sections) - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Immunohistochemistry (Frozen sections) - Anti-CD276 antibody [EPNCIR122] (ab134161)Image from Seaman S et al. Cancer Cell, 31, 501-515. Fig2.D doi:10.1016/j.ccell.2017.03.005. with permission from Elsevier

        Immunohistochemistry of mouse MC38 colon liver metastasis. Staining CD276 with ab134161 (green). Normal tissue (N)/tumor tissue (T) margins are indicated by a white dash.

         

      References

      This product has been referenced in:

      • Du H  et al. Antitumor Responses in the Absence of Toxicity in Solid Tumors by Targeting B7-H3 via Chimeric Antigen Receptor T Cells. Cancer Cell 35:221-237.e8 (2019). Read more (PubMed: 30753824) »
      • Xu Z  et al. High expression of B7-H3 and CD163 in cancer tissues indicates malignant clinicopathological status and poor prognosis of patients with urothelial cell carcinoma of the bladder. Oncol Lett 15:6519-6526 (2018). Read more (PubMed: 29725402) »
      See all 7 Publications for this product

      Publishing research using ab134161? Please let us know so that we can cite the reference in this datasheet.

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