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Active Cluster Of Differentiation 276 (CD276)

B7-H3; B7H3; CD276 Molecule; B7 homolog 3; Costimulatory molecule

ACTIVITY TEST

Cluster Of Differentiation 276 (CD276) also known as B7-H3 in human, is a member of the B7/CD28 superfamily of costimulatory molecules serving as an accessory modulator of T-cell response. 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. CD276 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. It has also been found to enhance the induction of primary cytotoxic T lymphocytes and stimulate IFN-gamma production. Besides, Sialic acid-binding Ig-like lectin 12 (SIGLEC12) has been identified as an interactor of CD276, thus a binding ELISA assay was conducted to detect the interaction of recombinant human CD276 and recombinant human SIGLEC12. Briefly, CD276 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to SIGLEC12-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-CD276 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of CD276 and SIGLEC12 was shown in Figure 1, and this effect was in a dose dependent manner.

USAGE

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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