Human IGSF8/CD316 Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB3117-100
MAB3117-SP
Detection of Human IGSF8/CD316 by Western Blot.
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Human IGSF8/CD316 Antibody Summary

Specificity
Detects human IGSF8/CD316 in direct ELISAs.
Source
Monoclonal Rabbit IgG Clone # 2587A
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
See below
Simple Western
20 µg/mL
Human Brain (Cerebellum)
Flow Cytometry
0.25 µg/106 cells
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 

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 Detection of Human IGSF8/CD316 antibody by Western Blot. View Larger

Detection of Human IGSF8/CD316 by Western Blot. Western blot shows lysates of TT human medullary thyroid cancer cell line and human brain (cerebellum) tissue. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human IGSF8/CD316 Monoclonal Antibody (Catalog # MAB3117) 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 Immunoblot Buffer Group 1.

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

Detection of IGSF8 in SHSY-5Y human cell line by Flow Cytometry. SHSY-5Y human neuroblastoma cell line was stained with Rabbit Anti-Human IGSF8 Monoclonal Antibody (Catalog # MAB3117, filled histogram) or Rabbit IgG isotype control antibody (Catalog # MAB1050, open histogram) followed by Anti-Rabbit IgG APC-conjugated Secondary Antibody (F0111). View our protocol for Staining Membrane-associated Proteins.

Simple Western View Larger

Detection of Human IGSF8/CD316 by Simple WesternTM. 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 IGSF8/CD316 Monoclonal Antibody (Catalog # MAB3117). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

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
  1. Clark, K.L. et al. (2001) J. Immunol. 167:5115.
  2. Stipp, C.S. et al. (2001) J. Biol. Chem. 276:40545.
  3. Charrin, S. et al. (2003) Biochem. J. 373:409.
  4. Stipp, C.S. et al. (2003) J. Cell Biol. 163:1167.
  5. Kettner, S. et al. (2007) Mol Cell Biol. 27:7718.
  6. Gordón-Alonso, M. et al. (2012) J Immunol 189:689.
  7. 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

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