Anti-CD276 antibody [RM2041] KO Tested (ab315907)

Knockout Tested Rabbit Recombinant Multiclonal CD276 antibody. Suitable for WB, IP, ICC/IF, Flow Cyt, IHC-P and reacts with Human samples.

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Images

Flow Cytometry - Anti-CD276 antibody [RM2041] (AB315907), expandable thumbnail

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: Multiclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IP	ICC/IF	Flow Cyt	IHC-Fr	IHC-P
Human	Tested	Tested	Tested	Tested	Not recommended	Tested
Mouse	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/5000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/200	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Associated Products

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1 product for Alternative Version

Target data

See full target information Cd276

Function

Modulates T-cell-mediated immune responses and the development of acute and chronic transplant rejection. Plays a positive regulatory role in bone formation and has a dual role in the bone-immune interface. Induces antitumor immunity as it activates both acquired and innate immunity leading to natural killer cell and CD8 T-cell dependent killing of tumor cells.

Alternative names

CD276, B7h3, Cd276, CD276 antigen, B7 homolog 3, Costimulatory molecule, B7-H3

Recommended products

Knockout Tested Rabbit Recombinant Multiclonal CD276 antibody. Suitable for WB, IP, ICC/IF, Flow Cyt, IHC-P and reacts with Human samples.

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: Multiclonal
Immunogens: The exact immunogen used to generate this antibody is proprietary information.
Clone number: RM2041
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

This product is a recombinant multiclonal 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.

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

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:
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10 product images

Flow Cytometry - Anti-CD276 antibody [RM2041] (ab315907)
Flow cytometric analysis of CD276 KO HEK293T (human CD276 knockout embryonic kidney epithelial cell, Left) / Parental HEK293T(Right) cells labelling CD276 with ab315907 at 1/5000 dilution (0.01 ug)/Red compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Immunoprecipitation - Anti-CD276 antibody [RM2041] (ab315907)
CD276 was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) whole cell lysate with ab315907 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315907 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: 293T (human embryonic kidney epithelial cell) whole cell lysate

Lane 2: ab315907 IP in 293T (human embryonic kidney epithelial cell) whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab315907 in 293T whole cell lysate
All lanes: Immunoprecipitation - Anti-CD276 antibody [RM2041] (ab315907) at 1/30 dilution
All lanes: 293T (human embryonic kidney epithelial cell) whole cell lysate with 5% NFDM/TBST
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution
Exposure time: 84s
Western blot - Anti-CD276 antibody [RM2041] (ab315907)
Low expression: skeletal muscle(PMID:36756390, PMID:35476450)
Samples are non-boiled as boiling may cause protein aggregation.
The identity of the lower MW bands in lane 1 and 2 are unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-CD276 antibody [RM2041] (ab315907) at 1/1000 dilution
Lane 1: Human lung tissue lysate at 20 µg with 5% NFDM/TBST
Lane 2: Human placenta tissue lysate at 20 µg with 5% NFDM/TBST
Lane 3: Human skeletal muscle tissue lysate at 20 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Observed band size: 90 kDa, 36 kDa
Exposure time: 3s
Immunocytochemistry/ Immunofluorescence - Anti-CD276 antibody [RM2041] (ab315907)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CD276 KO HEK293T (CD276 knockout human embryonic kidney epithelial cell) cells labelling CD276 with ab315907 at 1/100 (5.01 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).
Confocal image showing membranous staining in parental HEK293T cells, and no staining in CD276 KO HEK293T cells. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
Western blot - Anti-CD276 antibody [RM2041] (ab315907)
Lysates were freshly made, non-boiled and used immediately to minimize protein degradation.
All lanes: Western blot - Anti-CD276 antibody [RM2041] (ab315907) at 1/1000 dilution
Lane 1: HeLa (human epithelial cell line from cervical adenocarcinoma) whole cell lysate at 20 µg with 5% NFDM/TBST
Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 90 kDa
Exposure time: 37s
Western blot - Anti-CD276 antibody [RM2041] (ab315907)
In Western blot, ab315907 was shown to bind specifically to CD276.
Target of interest was observed at 90 KD in wild-type HEK293T cell lysates with no signal observed at this size in CD276 knockout cell line (lane 2, knockout cell line Human CD276 knockout HEK-293T cell line ab266658/ knockout cell lysate Human CD276 knockout HEK-293T cell lysate ab257097).
Lysates were freshly made, non-boiled and used immediately to minimize protein degradation.
In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842) staining at 1/100000 dilution.
All lanes: Western blot - Anti-CD276 antibody [RM2041] (ab315907) at 1/1000 dilution
Lane 1: Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST
Lane 2: CD276 knockout HEK293T whole cell lysate at 20 µg with 5% NFDM/TBST
Lane 3: LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST
Lane 4: Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Performed under reducing conditions.
Observed band size: 90 kDa, 15 kDa
Exposure time: 37s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [RM2041] (ab315907)
Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling CD276 with ab315907 at 1/200 (2.505 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Low expression: no staining on human skeletal muscle. The section was incubated with ab315907 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [RM2041] (ab315907)
Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling CD276 with ab315907 at 1/200 (2.505 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: no staining on human cardiac muscle. The section was incubated with ab315907 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [RM2041] (ab315907)
Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling CD276 with ab315907 at 1/200 (2.505 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human breast carcinoma. The section was incubated with ab315907 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [RM2041] (ab315907)
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CD276 with ab315907 at 1/200 (2.505 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human tonsil. The section was incubated with ab315907 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins