Human/Mouse/Rat IGSF8/CD316 Antibody

Catalog #: MAB31171 Datasheet / COA / SDS

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB31171-100
MAB31171-SP

Detection of Human, Mouse, and Rat IGSF8/CD316 by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse/Rat IGSF8/CD316 Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human IGSF8/CD316 in direct ELISAs. Detects human, mouse, and rat IGSF8/CD316 in Western blots.

Source

Recombinant Monoclonal Rabbit IgG Clone # 2587G

Purification

Protein A or G purified from cell culture supernatant

Immunogen

Human embryonic kidney cell, HEK293-derived human IGSF8/CD316
Arg28-Thr579
Accession # Q969P0-1

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

2 µg/mL

Human brain (hippocampus) tissue, Mouse brain (cerebellum) tissue, and Rat brain (cerebellum) tissue

Simple Western

20 µg/mL

Human Brain (Cerebellum)

CyTOF-ready

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

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells

SHSY-5Y human neuroblastoma cell line

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human, Mouse, and Rat IGSF8/CD316 by Western Blot. Western blot shows lysates of human brain (hippocampus) tissue, mouse brain (cerebellum) tissue, and rat brain (cerebellum) tissue. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human/Mouse/Rat IGSF8/CD316 Monoclonal Antibody (Catalog # MAB31171) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for IGSF8/CD316 at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Intracellular Staining by Flow Cytometry

Detection of IGSF8 antibody in SHSY-5Y human cell line antibody by Flow Cytometry.

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Detection of IGSF8 in SHSY-5Y human cell line by Flow Cytometry. SHSY-5Y human neuroblastoma cell line was stained with Rabbit Anti-Human/Mouse/Rat IGSF8/CD316 Monoclonal Antibody (Catalog # MAB31171, filled histogram) or Rabbit IgG isotype control antibody (MAB1050, open histogram) followed by Anti-Rabbit IgG APC-conjugated Secondary Antibody (F0111). Staining was performed using our Staining Membrane-associated Proteins protocol.

Simple Western

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Detection of Human/Mouse/Rat IGSF8/CD316 by Simple Western^TM. Simple Western lane view shows lysates of Human Brain (Cerebellum), loaded at 0.2 mg/mL. A specific band was detected for IGSF8/CD316 at approximately 73 kDa (as indicated) using 20 µg/mL of Rabbit Anti-Human/Mouse/Rat IGSF8/CD316 Monoclonal Antibody (Catalog # MAB31171). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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: IGSF8/CD316

IGSF8 (Immunoglobulin superfamily member 8), also known as EWI-2, KCT-4, LIR-D1, and PGRL, is a 75-kDa cell surface protein belonging to the immunoglobulin superfamily (1). IGSF8 is widely expressed, with pronounced mRNA expression in the brain and protein expression on peripheral blood lymphocytes and hepatocytes where it colocalizes with CD81 (1-3). It strongly associates with tetraspanins CD9 and CD81 which may act as physical linkers to form a complex with alpha 3 beta 1 integrin that may regulate cell aggregation and motility on laminin-5 (4). Human IGSF8 is synthesized as a 613 aa protein that includes a 27 aa signal peptide, a 552 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 13 aa cytoplasmic tail. Within the ECD, human IGSF8 shares 91% and 90% aa sequence identity with mouse and rat IGSF8, respectively. IGSF8 is an inducible receptor for Heat Shock Protein A8 (HSPA8) on activated dendritic cells (5). IGSF8 can interact with alpha -Actinin to regulate T cell immune synapses and HIV viral infection (6). In human glioma patients, low IGSF8 expression correlates with shorter survival time. Studies have shown that re-expression of IGSF8 in malignant glioblastoma cell lines inhibited glioblastoma colony formation in soft agar and caused diminished cell motility and invasion (7).

References

Clark, K.L. et al. (2001) J. Immunol. 167:5115.
Stipp, C.S. et al. (2001) J. Biol. Chem. 276:40545.
Charrin, S. et al. (2003) Biochem. J. 373:409.
Stipp, C.S. et al. (2003) J. Cell Biol. 163:1167.
Kettner, S. et al. (2007) Mol Cell Biol. 27:7718.
Gordón-Alonso, M. et al. (2012) J Immunol 189:689.
Kolesnikova, T. et al. (2009) Neoplasia.11:77.

Long Name

Immunoglobulin Superfamily, Member 8

Entrez Gene IDs

93185 (Human); 140559 (Mouse)

Alternate Names

CD316 antigen; CD316; CD81P3; CD81P3LIR-D1; EWI2; EWI-2; EWI2CD81 partner 3; Glu-Trp-Ile EWI motif-containing protein 2; IGSF8; immunoglobulin superfamily, member 8; KCT4; KCT-4; Keratinocytes-associated transmembrane protein 4; PGRL; PGRLimmunoglobulin superfamily member 8