Name: Anti-CD276 antibody [EPR20115]
SKU: ab219648
Rating: 4 (1 reviews)

Anti-CD276 antibody [EPR20115] (ab219648) is a rabbit monoclonal antibody detecting CD276 in Western Blot, IHC-P. Suitable for Human.

- KO validated for confirmed specificity

- Biophysical QC for unrivalled batch-batch consistency

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (AB219648), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

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

Abcam Recommends

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P
Human	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/4000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

See full target information CD276

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

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR20115
Purification technique: Affinity purification Protein A
Concentration

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

What is this antibody validated in?

Anti-CD276 antibody [EPR20115] (ab219648) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Immunohistochemistry (IHC-P) in Human samples.

What is the molecular weight of CD276?

Anti-CD276 [EPR20115] (ab219648) specifically detects a band for CD276 (UniProt: Q5ZPR3) at a molecular weight of 57kDa.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Specificity confirmed

The specificity of Anti-CD276 antibody [EPR20115] (ab219648) has been confirmed by Western blot testing in CD276 Knockout HEK293T cell line, ab266658.

Other related products

We have a range of other formats of antibody clone [EPR20115] also available for your convenience:
ab219648, Carrier free - ab227577, Carrier free - ab302848

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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.

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.

10 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (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.
Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
Lanes 1-4: Merged signal (red and green). Green - ab219648 observed at 90-110 kDa. Red - loading control Anti-GAPDH antibody [6C5] - 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 Human CD276 knockout HEK-293T cell line ab266658 (knockout cell lysate Human CD276 knockout HEK-293T cell lysate ab257097) was used. Wild-type and CD276 knockout samples were subjected to SDS-PAGE. ab219648 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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.
All lanes: Western blot - Anti-CD276 antibody [EPR20115] (ab219648) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: CD276 knockout HEK293T cell lysate at 20 µg
Lane 2: Western blot - Human CD276 knockout HEK-293T cell line (Human CD276 knockout HEK-293T cell line ab266658)
Lane 3: LNCaP cell lysate at 20 µg
Lane 4: Raji cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (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.
Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
Blocking/Dilution buffer: 5% NFDM/TBST.
Raji cells were used as a negative control for CD276 (B7-H3) (PMID: 21344157).
Negative control: lane 2 Raji cell lysate
All lanes: Western blot - Anti-CD276 antibody [EPR20115] (ab219648) at 1/1000 dilution
Lane 1: LNCaP (Human prostate cancer cell line) whole cell lysate at 10 µg
Lane 2: Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 57 kDa
Observed band size: 90-110 kDa
Exposure time: 3s
Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
Blocking/Dilution buffer: 5% NFDM/TBST.
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 [EPR20115] (ab219648) at 1/2000 dilution
Lane 1: U-2OS (Human bone osteosarcoma epithelial cell line) whole cell lysate at 10 µg
Lane 2: PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate at 10 µg
Lane 3: 293T (Human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg
Lane 4: MDA-MB-435S (Human ductal carcinoma cell line) whole cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 57 kDa
Observed band size: 90-110 kDa
Exposure time: 3s
Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2: 30 seconds; Lane 3: 10 seconds.
All lanes: Western blot - Anti-CD276 antibody [EPR20115] (ab219648) at 1/1000 dilution
Lane 1: Human lung lysate at 10 µg
Lane 2: Human skin lysate at 10 µg
Lane 3: Human placenta lysate at 10 µg
Secondary
All lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/4000 dilution
Predicted band size: 57 kDa
Observed band size: 90-110 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
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.
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] (ab219648)
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.
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] (ab219648)
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.
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] (ab219648)
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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.