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Anti-CD276 antibody [EPNCIR122] ab134161 is a rabbit monoclonal antibody that is used in CD276 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human and mouse samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with CD276 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- One antibody for all your CD276 staining, use in CD276 western blotting, IHC, immunofluorescence and flow cytometry

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Images

Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (AB134161), expandable thumbnail
  • Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (AB134161), expandable thumbnail
  • Western blot - Anti-CD276 antibody [EPNCIR122] (AB134161), expandable thumbnail
  • Immunoprecipitation - Anti-CD276 antibody [EPNCIR122] (AB134161), expandable thumbnail
  • Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (AB134161), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPFlow CytWBIHC-PICC/IFIHC-Fr
Human
Tested
Tested
Tested
Not recommended
Tested
Expected
Mouse
Expected
Expected
Expected
Not recommended
Expected
Tested

Tested
Tested

Species

Human

Dilution info

1/10 - 1/100

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000 - 1/10000

Notes

Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/200

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Mouse, Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1 µg/mL

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

-

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

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Target data

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.

Alternative names

B7H3, PSEC0249, UNQ309/PRO352, CD276, B7H3, PSEC0249, UNQ309/PRO352, CD276 antigen, 4Ig-B7-H3, B7 homolog 3, Costimulatory molecule, B7-H3

Recommended products

Anti-CD276 antibody [EPNCIR122] ab134161 is a rabbit monoclonal antibody that is used in CD276 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human and mouse samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with CD276 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- One antibody for all your CD276 staining, use in CD276 western blotting, IHC, immunofluorescence and flow cytometry

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPNCIR122

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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.

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 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 batch production

For more information, read more on recombinant antibodies.

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

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CD276 also known as B7-H3 is an immunoregulatory protein with a mass of approximately 57 kDa although post-translational modifications can increase its observed size. It is a member of the B7 family which is part of the immunoglobulin superfamily. CD276 is widely expressed in various tissues including placenta liver heart and also in many tumor cells. It is observed in lesser amounts in normal tissues making it an interesting target for cancer research. Various assays like HEK293 and HEK293T cell confluency studies often use CD276 to explore its expression under different conditions and seeding densities which are represented in confluency charts.

Biological function summary

CD276 modulates immune responses by acting as a co-inhibitory ligand impacting T-cell proliferation and cytokine synthesis. Not part of a known complex it plays a role in the adaptive immune system by providing negative feedback regulation that can inhibit the immune response. This capability has implications in preventing autoimmunity but may also contribute to tumor immune evasion by reducing effective anti-tumor responses. The B7-H3/CD276 interaction with receptors such as IL receptor family members suggests its diverse immunomodulatory functions.

Pathways

CD276 participates in the T-cell receptor (TCR) signaling pathways influencing immune checkpoint pathways. It interacts with other immune checkpoint proteins like PD-L1 and CTLA-4 but its exact receptor partner remains unidentified. The presence of CD276 in these pathways highlights its role in controlling T-cell activation and tolerance. By modulating TCR signals and impacting downstream effects CD276 serves as a regulatory point that can balance immune activation and inhibition critical in immune-mediated conditions.

Associated diseases and disorders

CD276 has strong associations with cancer particularly in solid tumors such as prostate and breast cancer. It is often overexpressed in malignant tissues contributing to immune evasion by tumors. Researchers propose it as a potential target for cancer immunotherapy due to this attribute. CD276's link to disorders like asthma has also been suggested where its interaction with proteins involved in inflammatory pathways such as IL response elements can exacerbate the condition. This implicates it as a dual-role protein in both disease progression and potential therapeutic targeting.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

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

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

    Flow cytometry overlay histogram showing wild-type HEK293 (green line) and CD276 knockout HEK293 (Human CD276 knockout HEK-293T cell line ab266658) cells stained with ab134161 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab134161) (1x106 in 100μl at 0.2 μg/ml) for 30 min at 4°C.

    The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) was used at 1/2000 for 30 min at 4°C.

    Isotype control antibody was Rabbit IgG (monoclonal) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) used at the same concentration and conditions as the primary antibody (wild-type HEK293 - black line CD276 HEK293 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

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

    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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution. Isotype control antibody (black line) was rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Performed using purified antibody.

  • Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161), expandable thumbnail

    Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)

    We recommend not to boil the samples after lysis to get desired WB results.

    CD276 (B7-H3) has a molecular weight of about 90–100 kDa, depending on the glycosylation levels. [PMID:22473715].

    All lanes: Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161) at 1/1000 dilution

    Lane 1: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate boiled. at 20 µg

    Lane 2: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate unboiled. at 20 µg

    Lane 3: LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate boiled. at 20 µg

    Lane 4: LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate unboiled. at 20 µg

    Secondary

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

    Predicted band size: 57 kDa

    Observed band size: 90 kDa

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

    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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab134161 in HEK 293 whole cell lysate.

    Blocking/dilution buffer: 5% NFDM/TBST

    Performed using purified antibody.

    All lanes: Immunoprecipitation - Anti-CD276 antibody [EPNCIR122] (ab134161)

    Predicted band size: 57 kDa

    Exposure time: 3s

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

    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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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), expandable thumbnail

    Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)

    Blocking/dilution buffer: 5% NFDM/TBST

    Performed using purified antibody.

    All lanes: Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161) at 1/200 dilution

    All lanes: CHO (hamster ovary epithelial cell) whole cell lysates transfected with human CD276 at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution

    Predicted band size: 57 kDa

    Observed band size: 110 kDa

    Exposure time: 3min

  • Immunocytochemistry/ Immunofluorescence - Anti-CD276 antibody [EPNCIR122] (ab134161), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-CD276 antibody [EPNCIR122] (ab134161)

    ab134161 staining CD276 in wild-type HEK293 cells (top panel) and CD276 knockout HEK293 cells (Human CD276 knockout HEK-293T cell line ab266658) (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab134161 at 1μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

  • Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161), expandable thumbnail

    Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)

    Blocking/dilution buffer: 5% NFDM/TBST

    Performed using purified antibody.

    All lanes: Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161) at 1/1000 dilution

    All lanes: LNCap (human prostate carcinoma epithelial cell) whole cell lysate at 15 µg

    Secondary

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

    Predicted band size: 112 kDa, 388 kDa, 47 kDa, 55 kDa, 57 kDa, 72 kDa, 83 kDa, 97 kDa

    Observed band size: 100 kDa, 110 kDa, 140 kDa, 388 kDa, 50-70 kDa

    Exposure time: 3min

  • Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161), expandable thumbnail

    Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)

    Blocking/dilution buffer: 5% NFDM/TBST

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

    Lane 1: CHO-K1 cell lysates, transfected with Human CD276 at 20 µg

    Lane 2: CHO-K1 cell lysates, transfected with Mouse CD276. at 20 µg

    Secondary

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

    Predicted band size: 57 kDa

    Exposure time: 3min

  • Immunohistochemistry (Frozen sections) - Anti-CD276 antibody [EPNCIR122] (ab134161), expandable thumbnail
    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 (Frozen sections) - Anti-CD276 antibody [EPNCIR122] (ab134161)

    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.

  • Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161), expandable thumbnail

    Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)

    Blocking and dilution buffer: 5% NFDM/TBST.

    We recommend not to boil the samples after lysis to get desired WB results.

    Mouse CD276 (B7-H3) has a molecular weight about 45–66 kDa, depending on the glycosylation levels [PMID: 34366684].

    All lanes: Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161) at 1/1000 dilution

    Lane 1: LLC (Mouse Lewis lung carcinoma cell) whole cell lysate (boiled) at 20 µg

    Lane 2: LLC (Mouse Lewis lung carcinoma cell) whole cell lysate (unboiled) at 20 µg

    Secondary

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

    Predicted band size: 57 kDa

    Observed band size: 45-66 kDa

    Exposure time: 180s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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