This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?

Name: Recombinant Human B7-H3 His-tag Protein
Brand: R&D Systems
SKU: 1949-B3
Price: 405 USD

The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.

Recombinant Human B7-H3 His-tag Protein

Catalog # 1949-B3 | R&D Systems, Inc. a Bio-Techne Brand

Citations (4)

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Key Product Details

Source:	NS0
Accession #:	NP_079516
Applications:	Bioactivity

View Full Technical Details for Recombinant Human B7-H3 His-tag Protein

Product Specifications for Recombinant Human B7-H3 His-tag Protein

Source	Mouse myeloma cell line, NS0-derived human B7-H3 protein Leu29-Pro245, with a C-terminal 10-His tag
Purity	>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin level	<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Leu29
Predicted Molecular Mass	24.8 kDa
SDS-PAGE	38-48 kDa, reducing conditions
Activity	Measured in a cell proliferation assay using mouse CD4⁺ cells in the presence of anti-CD3. The ED₅₀ for this effect is 3-12 µg/mL.

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

1949-B3

Carrier

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Reconstitution	Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

1949-B3/CF

Carrier Free

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 100 μg/mL in sterile PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: B7-H3

T cells require a signal induced by the engagement of the T cell receptor and a “co-stimulatory” signal(s) through distinct T cell surface molecules for optimal T cell expansion and activation. Members of the B7 superfamily of counter-receptors were identified by their ability to interact with co-stimulatory molecules found on the surface of T cells. Members of the B7 superfamily include B7-1 (CD80), B7-2 (CD86), B7-H1 (PD-L1), B7-H2 (B7RP-1), PD-L2 (B7-DC), and B7-H3 (1). Human B7-H3 was originally identified as a 316 amino acid (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 (2). Subsequently, a second dominantly expressed form of human B7-H3, containing a splice variation that duplicates the V-like and C-like Ig domains, was found. The 534 aa splice variant of B7-H3 has been referred to as B7-H3b, 4Ig-B7-H3, and B7-H3VCVC (3 - 5). RT-PCR transcripts for both B7-H3 and 4Ig‑B7‑H3 have been found in all tissues except peripheral blood leukocytes (5). Southern blot analysis indicates that the 4Ig-B7-H3 isoform of B7‑H3 is the predominate isoform expressed in human tissues (5). In mouse, only a single form of B7-H3 containing one V-like and one C-like Ig domain was detected (5). Mouse and human B7-H3 share 87% aa sequence identity (3). B7-H3 has been shown to be expressed at very high levels in immature dendritic cells, at moderate levels on mature dendritic cells, LPS stimulated immature dendritic cells and LPS stimulated monocytes, and at low levels on resting monocytes (3). B7-H3 binds to activated T cells via an as yet unidentified receptor. In assays using sub-optimal amount so anti-CD3 stimulation, 2Ig‑B7‑H3 enhances T cell proliferation, T cell interferon-gamma (IFN-gamma ) production, and cytotoxic T cells induction (2). In an assay in which cells were transfected with 4Ig-B7-H3 or 2Ig‑B7‑H3 and anti-CD3 antibody was added, neither 4Ig-B7-H3 nor 2Ig‑B7‑H3 was capable of co‑stimulating T cell proliferation or IFN-gamma production (4, 5).

Item	Value
References	Coyle, A.J. and J.-C. Gutierrez-Ramos (2001) Nature Immunol. 2:203. Chapoval, A.I. et al. (2001) Nature Immunol. 2:269. Sun, M. et al. (2002) J. Immunol. 168:6294. Steinberger, et al. (2004) J. Immunol. 172:2352. Ling, V. et al. (2003) Genomic 82:365.
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 Recombinant Human B7-H3 His-tag Protein (4)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant Human B7-H3 His-tag Protein include:

Species: Human
Sample Types: Recombinant Protein
Applications: ELISA (Capture)

IJG Burvenich et al. (2018-07-30), Molecular imaging of T cell co-regulator factor B7-H3 with 89Zr-DS-5573a Theranostics, 2018-08(15):4199-4209.
PMID: 30128047
Sample Types: Protein
Applications: Bioassay

D Quandt et al. (2017-04-11), High constitutive B7-H3 expression on human keratinocytes supports T cell immunity J. Dermatol. Sci., 2017-00(0).
PMID: 28465042
Species: Human
Sample Types: Whole Tissue
Applications: Bioassay

Ingebrigtsen V et al. (2014-08-20), B7-H3 expression in colorectal cancer: associations with clinicopathological parameters and patient outcome. BMC Cancer, 2014-014(0):602.
PMID: 25139714
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

Zhang G et al. (2010-08-09), B7-h3 augments the inflammatory response and is associated with human sepsis. J. Immunol., 2010-0185(6):3677-84.
PMID: 20696859

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Showing 1 - 4 of 4 citations Showing All

FAQs for Recombinant Human B7-H3 His-tag Protein

This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?

The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.

Recombinant Human B7-H3 His-tag Protein

Catalog # 1949-B3 | R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

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Product Specifications for Recombinant Human B7-H3 His-tag Protein

Formulation, Preparation and Storage

Carrier Free

1949-B3

Carrier

1949-B3/CF

Carrier Free

Reconstitution Calculator

Background: B7-H3

Citations for Recombinant Human B7-H3 His-tag Protein (4)

FAQs for Recombinant Human B7-H3 His-tag Protein

Product Documents for Recombinant Human B7-H3 His-tag Protein

Product Datasheets

COA

Certificate of Analysis

SDS

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