Western blot shows lysates of HEK293T human embryonic kidney cell line either mock transfected or transfected with human VSIG4. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human VSIG4 Monoclonal Antibody (Catalog # MAB4646) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for VSIG4 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of VSIG4 in Human Macrophages by Flow Cytometry

Human CD14+ monocytes were positively selected from PBMC (MAGH105) and cultured for 7 days with 10% Human AB serum, 50 ng/ml M-CSF (216-MC), and 10μM hydrocortisone (4093). Cells were harvested and stained with either (A) Mouse anti-Human VSIG4 (Catalog # MAB4646) or (B) Isotype control antibody (MAB003) followed by Allophycocyanin-conjugated anti-Mouse IgG Secondary Antibody (F0101B) and Mouse anti-Human CD11b Alexa Fluor® 488-conjugated Monoclonal Antibody (FAB16991G). Staining was performed using our Staining Membrane-Associated Proteins protocol.

Applications for Human VSIG4 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: Human PBMC-derived macrophages

Western Blot

2 µg/mL
Sample: HEK293T human embryonic kidney cell line transfected with human VSIG4

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: VSIG4

VSIG4 (Vset and immunoglobulin domain containing 4), also known as complement receptor immunoglobulin (CRIg) and Z39IG, is a 45 kDa, type I transmembrane protein of the B7 family within the Ig superfamily that is expressed only in tissue resident macrophages (1-4). The gene is located on the X chromosome (2). The human VSIG4 cDNA encodes 399 amino acids (aa) including a 19 aa signal sequence, a 264 aa extracellular domain (ECD) containing a V-type and a C2-type Ig domain, a 21 aa transmembrane domain and a 95 aa cytoplasmic domain (3). The human VSIG4 ECD shares 84% aa identity with canine VSIG4. Within the IgV domain, it shares 90%, 80% and 78% aa identity with bovine, mouse and rat VSIG4, respectively; these animals lack the C2-type domain. Splice isoforms of 321, 305, 272, 201 and 199 aa lack all or part of the cytoplasmic domain, the C2-type Ig domain and/or the transmembrane domain (5). VSIG4 is specifically expressed on macrophages in the thymic medulla, peritoneum, alveoli, synovia, adipose and heart, liver Kupffer cells, placental Hofbauer cells, and atherosclerotic foam cells (1-4, 6-9). It is absent on infiltrating macrophages (8). VSIG4 is a complement receptor that binds C3b and iC3b fragments, internalizes them to recycling endosomes, and is recycled to the cell surface (4, 6). It contributes significantly to innate immunity by binding and phagocytosis of complement opsonized invading pathogens (4, 8, 10). Binding of either native or recombinant soluble VSIG4 to C3b inhibits complement amplification through the alternative, but not classical, pathway (10, 11). VSIG4 is also a negative regulator of mouse and human T cell activation (2). Although VSIG4 engagement may activate NF kappaB and thus be proinflammatory in some cases, many of its activities are important in resolving, rather than initiating, inflammation (1, 2, 7, 10, 11). There is emerging evidence in human conditions that VSIG4 may be a valuable biomarker in infection and immunity, inflammatory conditions and cancer prognosis (12, 13, 14).

References

He, J.Q. et al. (2008) Mol. Immunol. 4041.
Vogt, L. et al. (2006) J. Clin. Invest. 116:2817.
Langnaese, K. et al. (2000) Biochim. Biophys. Acta 1492:522.
Helmy, K. et al. (2006) Cell 124:915.
Entrez protein Accession # EAX05393, NP_001093901, CAI42052, CAI4205, EAX05394.
Tanaka, M. et al. (2008) Clin. Exp. Immunol. 154:38.
Lee, M.Y. et al. (2006) J. Leukoc. Biol. 80:922.
Gorgani, N.N. et al. (2008) J. Immunol. 181:7902.
Walker, M.G. (2002) Biochim. Biophys. Acta 1574:387.
Wiesmann, C. et al. (2006) Nature 444:217.
Katschke, K.J. et al. (2007) J. Exp. Med. 204:1319.
Small, A.G. et al. (2016) Swiss Med Wkly. 5:146.
Roh J. et al. (2017) Oncotarget. 8:58122.
Kim K.H. et al. (2016) Autophagy. 12:1647.

Long Name

V-Set and Immunoglobulin Domain Containing 4

Alternate Names

Z39IG

Entrez Gene IDs

11326 (Human); 278180 (Mouse); 312102 (Rat); 102144784 (Cynomolgus Monkey)

Gene Symbol

VSIG4

UniProt

Q9Y279

Additional VSIG4 Products

Product Documents for Human VSIG4 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human VSIG4 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human VSIG4 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB4646

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

View all VSIG4 Antibodies »

Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human VSIG4 Antibody

Detection of Human VSIG4 by Western Blot.