Human B7-H3 MAb (Clone 185504)

Name: Human B7-H3 MAb (Clone 185504)
Brand: R&D Systems
SKU: MAB1027
Rating: 5 (4 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # MAB1027

Citations (8)

(4)

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Key Product Details

Species Reactivity:	Human
Immunogen:	Mouse myeloma cell line NS0-derived recombinant human B7‑H3 Leu29-Pro245 Accession # NP_079516
Applications:	CyTOF-ready, Flow Cytometry, Knockout Validated, Western Blot
Label:	Unconjugated
Source:	Monoclonal Mouse IgG₁ Clone # 185504
Specificity:	Detects human B7-H3 in direct ELISAs and Western blots. In Western blots, no cross-reactivity with recombinant human (rh) B7‑H1, rhB7‑H2, rhB7‑1, rhB7‑2, or recombinant mouse PD-L2 is observed.

View Full Technical Details for Human B7-H3 MAb (Clone 185504)

Applications for Human B7-H3 MAb (Clone 185504)

	Recommended Concentration	Sample
Western Blot	2 µg/mL	See below
Flow Cytometry	0.25 µg/10⁶ cells	See below
CyTOF-ready	Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Item	Value
Knockout Validated	B7‑H3 is specifically detected in U2OS human osteosarcoma parental cell line but is not detectable in B7‑H3 knockout U2OS cell line.

Please Note: Optimal dilutions should be determined by each laboratory for each application.

Scientific Data Examples for Human B7-H3 MAb (Clone 185504)

	Western Blot Detection of Human B7‑H3 by Western Blot. Western blot shows lysates of LNCaP human prostate cancer cell line and U2OS human osteosarcoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human B7-H3 Monoclonal Antibody (Catalog # MAB1027) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for B7-H3 at approximately 90-110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
	Flow Cytometry Detection of B7-H3 in PC‑3 Human Cell Line by Flow Cytometry. PC-3 human prostate cancer cell line was stained with Mouse Anti-Human B7-H3 Monoclonal Antibody (Catalog # MAB1027, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0102B).
	Flow Cytometry Detection of B7‑H3 in Human Dendritic Cells by Flow Cytometry. Human monocyte-derived dendritic cells were stained with Mouse Anti-Human B7-H3 Monoclonal Antibody (Catalog # MAB1027, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B).
	Knockout Validated Western Blot Shows Human B7‑H3 Specificity by Using Knockout Cell Line. Western blot shows lysates of U2OS human osteosarcoma parental cell line and B7-H3 knockout U2OS cell line (KO). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human B7-H3 Monoclonal Antibody (Catalog # MAB1027) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for B7-H3 at approximately 95 kDa (as indicated) in the parental U2OS cell line, but is not detectable in knockout U2OS cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Formulation, Preparation and Storage

Purification	Protein A or G purified from hybridoma culture supernatant
Reconstitution	Reconstitute at 0.5 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 6 months, -20 to -70 °C under sterile conditions after reconstitution.

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Background: B7-H3

Human B7 homolog 3 (B7-H3) is a member of the B7 family of immune proteins that provide signals for regulating immune responses (1‑3). Other family members include B7-1, B7-2, B7-H2, PD-L1 (B7-H1), and PD-L2. B7 proteins are immunoglobulin (Ig) superfamily members with extracellular Ig-V-like and Ig-C-like domains and short cytoplasmic domains. Among the family members, they share about 20‑40% amino acid (aa) sequence identity. The cloned human B7-H3 cDNA encodes a 316 aa type I membrane precursor protein with a putative 28 aa signal peptide, a 217 aa extracellular region containing one V-like and one C-like Ig domain, a transmembrane region, and a 45 aa cytoplasmic domain. An isoform of human B7-H3 containing a four-Ig-like domain extracellular region has also been identified. Human B7-H3 is not expressed on resting B cells, T cells, monocytes or dendritic cells, but is induced on dendritic cells and monocytes by inflammatory cytokines. B7-H3 expression is also detected on various normal tissues and in some tumor cell lines. Human B7-H3 does not bind any known members of the CD28 family of immunoreceptors. However, B7-H3 has been shown to bind an unidentified counter-receptor on activated T cells to costimulate the proliferation of CD4+ or CD8+ T cells. B7-H3 has also been found to enhance the induction of primary cytotoxic T lymphocytes and stimulate IFN-gamma production (1‑3).

Item	Value
References	Chapoval, A.I. et al. (2001) Nat. Immunol. 2:269. Sharpe, A.H. and G.J. Freeman (2002) Nat. Rev. Immunol. 2:116. Coyle, A. and J. Gutierrez-Ramos (2001) Nat. Immunol. 2:203.
Long Name	B7 Homolog 3
Entrez Gene IDs	80381 (Human); 102657 (Mouse); 315716 (Rat); 102131295 (Cynomolgus Monkey)
Alternate Names	B7-H3, B7-H3B7 homolog 3, B7H3, B7H34Ig-B7-H3, CD276, CD276 antigen, CD276 molecule, Costimulatory molecule

Citations for Human B7-H3 MAb (Clone 185504) (8)

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Citations are publications that use Bio-Techne products. Selected citations for Human B7-H3 MAb (Clone 185504) include:

Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

M Oroujeni et al. (2022), Evaluation of an Affibody-Based Binder for Imaging of Immune Check-Point Molecule B7-H3 Pharmaceutics, 14(9).
PMID: 36145529
Species: Human
Sample Types: Whole Tissue
Applications: IHC

J Liu et al. (2021), Targeting B7-H3 via chimeric antigen receptor T cells and bispecific killer cell engagers augments antitumor response of cytotoxic lymphocytes Journal of hematology oncology, 2021;14(1):21.
PMID: 33514401
Species: Human
Sample Types: Cell Lysates
Applications: Immunoprecipitation

S Yang et al. (2020), Targeting B7-H3 Immune Checkpoint With Chimeric Antigen Receptor-Engineered Natural Killer Cells Exhibits Potent Cytotoxicity Against Non-Small Cell Lung Cancer Front Pharmacol, 11(0):1089.
PMID: 32848731
Species: Human
Sample Types: Cell Lysates
Applications: Immunoprecipitation

S Yang et al. (2020), Targeting B7-H3 Immune Checkpoint With Chimeric Antigen Receptor-Engineered Natural Killer Cells Exhibits Potent Cytotoxicity Against Non-Small Cell Lung Cancer Front Pharmacol, 11(0):1089.
PMID: 32848731
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

Sucker A et al. (2014), Genetic evolution of T-cell resistance in the course of melanoma progression. Clin Cancer Res, 20(24):6593-604.
PMID: 25294904
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

Ramsay et al. (2012), Multiple inhibitory ligands induce impaired T-cell immunologic synapse function in chronic lymphocytic leukemia that can be blocked with lenalidomide: establishing a reversible immune evasion mechanism in human cancer. Blood, 120(7):1412-21.
PMID: 22547582
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

Tran CN et al. (2008), Interactions of T cells with fibroblast-like synoviocytes: role of the B7 family costimulatory ligand B7-H3. J. Immunol., 180(5):2989-98.
PMID: 18292521
Species: Human
Sample Types: Cell Culture Supernates
Applications: Western Blot

Das S et al. (2006), Expression of B7-H1 on gastric epithelial cells: its potential role in regulating T cells during Helicobacter pylori infection. J. Immunol., 176(5):3000-9.
PMID: 16493058

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Customer Reviews for Human B7-H3 MAb (Clone 185504) (4)

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Human B7-H3 MAb (Clone 185504)

Name: Anonymous

Application: WB

Sample Tested: Osteosarcoma U2OS cell line

Species: Human

Verified Customer | Posted 06/22/2022
Human B7-H3 Antibody

Name: Anonymous

Application: Simple Western

Sample Tested: Purified protein

Species: Human

Verified Customer | Posted 05/01/2020
Human B7-H3 Antibody

Name: Jenna Nguyen

Application: Func

Sample Tested: 3T3-L1 mouse embryonic fibroblast adipose-like cell line

Species: Mouse

Verified Customer | Posted 05/07/2019
Human B7-H3 Antibody

Name: Anonymous

Application: Simple Western

Sample Tested: Purified protein

Species: Human

Verified Customer | Posted 11/02/2018

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Product Documents for Human B7-H3 MAb (Clone 185504)

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Human B7-H3 MAb (Clone 185504)

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Applications for Human B7-H3 MAb (Clone 185504)

Scientific Data Examples for Human B7-H3 MAb (Clone 185504)

Western Blot

Flow Cytometry

Flow Cytometry

Knockout Validated

Formulation, Preparation and Storage

Reconstitution Calculator

Background: B7-H3

Citations for Human B7-H3 MAb (Clone 185504) (8)

Customer Reviews for Human B7-H3 MAb (Clone 185504) (4)

Customer Images

Human B7-H3 MAb (Clone 185504)

Human B7-H3 Antibody

Human B7-H3 Antibody

Human B7-H3 Antibody

Product Documents for Human B7-H3 MAb (Clone 185504)

Product Datasheets

COA

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