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CD276 Recombinant Rabbit Monoclonal Antibody [PD00-36]

Usd: 385

Specification

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Catalog# HA721245

CD276 Recombinant Rabbit Monoclonal Antibody [PD00-36]

Application

  • IHC-P

  • FC

  • WB

  • IF-Tissue

Reactivity

  • Human

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

CD276 Recombinant Rabbit Monoclonal Antibody [PD00-36]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within C terminal human CD276.

Species Reactivity

Human

Validated Applications

IHC-P, FC, WB, IF-Tissue

Molecular Weight

Predicted band size: 57 kDa

Positive Control

HeLa cell lysate, MCF7 cell lysate, HEK-293 cell lysate, LoVo cell lysate, U-2 OS cell lysate, LNCaP cell lysate, SH-SY5Y cell lysate, THP-1 cell lysate, HCT 116 cell lysate, HS-SY-II cell lysate, human lung squamous cell carcinoma tissue, human lung adenocarcinoma tissue, human prostate carcinoma tissue, HEK-293, THP-1.

Conjugation

unconjugated

Clone Number

PD00-36

RRID

AB_3068338

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • IHC-P

  • 1:5,000

  • FC

  • 1:500-1:1,000

  • WB

  • 1:2,000-1:5,000

  • IF-Tissue

  • 1:1,000

Target

Function

The protein encoded by this gene belongs to the immunoglobulin superfamily. 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.

Background References

1. Zhou WT et al. B7-H3/CD276: An Emerging Cancer Immunotherapy. Front Immunol. 2021 Jul

2. Liu S et al. The Role of CD276 in Cancers. Front Oncol. 2021 Mar

Subcellular Location

Membrane.

UNIPROT

SwissProt: Q5ZPR3 Human

Synonyms

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

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of CD276 on different lysates with Rabbit anti-CD276 antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: MCF7 cell lysate (20 µg/Lane)<br />Lane 3: HEK-293 cell lysate (20 µg/Lane)<br />Lane 4: LoVo cell lysate (20 µg/Lane)<br />Lane 5: U-2 OS cell lysate (20 µg/Lane)<br />Lane 6: LNCaP cell lysate (20 µg/Lane)<br />Lane 7: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 8: THP-1 cell lysate (20 µg/Lane)<br />Lane 9: HCT 116 cell lysate (20 µg/Lane)<br />Lane 10: Raji cell lysate (negative) (20 µg/Lane)<br /><br />Predicted band size: 57 kDa<br />Observed band size: 100 kDa<br /><br />Exposure time: 6 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of CD276 on different lysates with Rabbit anti-CD276 antibody (HA721245) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: MCF7 cell lysate (20 µg/Lane)
    Lane 3: HEK-293 cell lysate (20 µg/Lane)
    Lane 4: LoVo cell lysate (20 µg/Lane)
    Lane 5: U-2 OS cell lysate (20 µg/Lane)
    Lane 6: LNCaP cell lysate (20 µg/Lane)
    Lane 7: SH-SY5Y cell lysate (20 µg/Lane)
    Lane 8: THP-1 cell lysate (20 µg/Lane)
    Lane 9: HCT 116 cell lysate (20 µg/Lane)
    Lane 10: Raji cell lysate (negative) (20 µg/Lane)

    Predicted band size: 57 kDa
    Observed band size: 100 kDa

    Exposure time: 6 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721245) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of CD276 on different lysates with Rabbit anti-CD276 antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>) at 1/5,000 dilution.<br /><br />Lane 1: HS-SY-II-si NT cell lysate<br />Lane 2: HS-SY-II-si CD276 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 57 kDa<br />Observed band size: 90 kDa<br /><br />Exposure time: 1 minute 2 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>) at 1/5,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of CD276 on different lysates with Rabbit anti-CD276 antibody (HA721245) at 1/5,000 dilution.

    Lane 1: HS-SY-II-si NT cell lysate
    Lane 2: HS-SY-II-si CD276 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 57 kDa
    Observed band size: 90 kDa

    Exposure time: 1 minute 2 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721245) at 1/5,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human lung squamous cell carcinoma tissue with Rabbit anti-CD276 antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human lung squamous cell carcinoma tissue with Rabbit anti-CD276 antibody (HA721245) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721245) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma tissue with Rabbit anti-CD276 antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma tissue with Rabbit anti-CD276 antibody (HA721245) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721245) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue with Rabbit anti-CD276 antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue with Rabbit anti-CD276 antibody (HA721245) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721245) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of HEK-293 cells labeling CD276.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HEK-293 cells labeling CD276.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721245, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Flow cytometric analysis of THP-1 cells labeling CD276.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721245" style="font-weight: bold;text-decoration: underline;">HA721245</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of THP-1 cells labeling CD276.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721245, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Application: IF-Tissue<br /><br />Species: Human<br /><br />Site: prostate carcinoma<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/1,000

    Application: IF-Tissue

    Species: Human

    Site: prostate carcinoma

    Sample: Paraffin-embedded section

    Antibody concentration: 1/1,000

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Breast carcinoma<br />Sample: Paraffin-embedded section<br /><br />Primary antibody dilution: 1/2,000<br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes<br />Platform: Leica Biosystems BOND&reg; RX

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Breast carcinoma
    Sample: Paraffin-embedded section

    Primary antibody dilution: 1/2,000
    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes
    Platform: Leica Biosystems BOND® RX

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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