CD276 Antibody [P14M24] | Primary Antibodies

Application: Reactivity: Mouse, Human

Usage Information

Dilution
WB1:200 IP1:10 - 1:100 IF1:1700 FCM1:1000-1:10000

Application
WB, IP, IHC, IF, FCM

Reactivity
Mouse, Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
388 kDa,47-112 kDa 388 kDa, 45-140 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight.

Positive Control	Mouse MC38 colon liver metastasis; THP1 cells; LNCap cells; HEK 293 cells; LLC cells
Negative Control	HEK-293 cells (boiled);LNCaP cells (boiled); LLC cells (boiled)

Datasheet & SDS

Biological Description

Specificity
CD276 Antibody [P14M24] detects endogenous levels of total CD276 protein.

Clone
P14M24

Synonym(s)
CD276; B7H3; PSEC0249; UNQ309/PRO352; CD276 antigen; 4Ig-B7-H3; B7 homolog 3; Costimulatory molecule; B7-H3

Background
CD276 (B7-H3) is a type I transmembrane glycoprotein of the B7 family in the immunoglobulin superfamily (~57 kDa core, post-translationally glycosylated and phosphorylated). In mice, CD276 exists as a single IgV-IgC ectodomain, while in humans it has a double IgV-IgC ectodomain structure. It primarily functions as a co-inhibitory immune checkpoint ligand, overexpressed in tumor cells and antigen-presenting cells (APCs) to suppress T-cell proliferation and IFNγ production via unidentified receptor(s) on activated T cells. CD276 contains a signal peptide, extracellular Ig-like domains (with an N-terminal IgV domain conferring ligand specificity and tandem IgC2-set domains stabilized by disulfide bonds), a short transmembrane helix, and a cytoplasmic tail. The cytoplasmic tail includes a Tyr288 phosphorylation site that recruits Src/Lyn kinases, palmitoylation motifs for lipid raft localization, and endocytic signals. Its core activity involves ligand-receptor engagement that inhibits T-cell responses (reducing IL-2 and IFNγ, increasing anergy), suppresses natural killer cell cytotoxicity, and facilitates tumor immune evasion. CD276 also promotes non-immune functions, such as enhancing glycolysis and oxidative phosphorylation in adipocyte progenitors via Myc network activation, thereby limiting lipid accumulation and obesity. CD276 regulates adaptive immunity through negative costimulation, modulates APC function, inhibits angiogenesis by mimicking thrombospondin-1, and maintains metabolic homeostasis in adipose tissue by restraining adipocyte precursor differentiation. Its basal expression is low and restricted, but it is overexpressed in more than 90% of human cancers, including prostate, lung, melanoma, and glioblastoma, where it correlates with poor prognosis due to immune escape and metastasis.

Background

CD276 (B7-H3) is a type I transmembrane glycoprotein of the B7 family in the immunoglobulin superfamily (~57 kDa core, post-translationally glycosylated and phosphorylated). In mice, CD276 exists as a single IgV-IgC ectodomain, while in humans it has a double IgV-IgC ectodomain structure. It primarily functions as a co-inhibitory immune checkpoint ligand, overexpressed in tumor cells and antigen-presenting cells (APCs) to suppress T-cell proliferation and IFNγ production via unidentified receptor(s) on activated T cells. CD276 contains a signal peptide, extracellular Ig-like domains (with an N-terminal IgV domain conferring ligand specificity and tandem IgC2-set domains stabilized by disulfide bonds), a short transmembrane helix, and a cytoplasmic tail. The cytoplasmic tail includes a Tyr288 phosphorylation site that recruits Src/Lyn kinases, palmitoylation motifs for lipid raft localization, and endocytic signals. Its core activity involves ligand-receptor engagement that inhibits T-cell responses (reducing IL-2 and IFNγ, increasing anergy), suppresses natural killer cell cytotoxicity, and facilitates tumor immune evasion. CD276 also promotes non-immune functions, such as enhancing glycolysis and oxidative phosphorylation in adipocyte progenitors via Myc network activation, thereby limiting lipid accumulation and obesity. CD276 regulates adaptive immunity through negative costimulation, modulates APC function, inhibits angiogenesis by mimicking thrombospondin-1, and maintains metabolic homeostasis in adipose tissue by restraining adipocyte precursor differentiation. Its basal expression is low and restricted, but it is overexpressed in more than 90% of human cancers, including prostate, lung, melanoma, and glioblastoma, where it correlates with poor prognosis due to immune escape and metastasis.

References
https://pubmed.ncbi.nlm.nih.gov/36859240/ https://pubmed.ncbi.nlm.nih.gov/40937367/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%