TSPAN8/TM4SF3 Antibody (458811) [Unconjugated]

Western blot shows lysates of COLO 205 human colorectal adenocarcinoma cell line and HT‑29 human colon adenocarcinoma cell line. PVDF membrane was probed with 2 µg/mL of Rat Anti-Human TSPAN8 Monoclonal Antibody (Catalog # MAB4734) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for TSPAN8 at approximately 35 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Immunoblot Buffer Group 1.

HT‑29 human colon adenocarcinoma cell line was stained with Rat Anti-Human TSPAN8 Monoclonal Antibody (Catalog # MAB4734, filled histogram) or isotype control antibody (Catalog # MAB0061, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG F(ab')2 Secondary Antibody (Catalog # F0113). View our protocol for Staining Membrane-associated Proteins.

TSPAN8 was detected in immersion fixed HT‑29 human colon adenocarcinoma cell line using Rat Anti-Human TSPAN8 Monoclonal Antibody (Catalog # MAB4734) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

TSPAN8 was detected in immersion fixed HT‑29 Human Colon Adenocarcinoma Cell Line (Positive) and Daudi Human Burkitt's Lymphoma Cell Line (Negative) Cells using Rat Anti-Human TSPAN8 Monoclonal Antibody (Catalog # MAB4734) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surface. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

• Reactivity: Hu
• Applications: WB, Flow, CyTOF-ready, ICC/IF
• Clone: 458811
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Concentration: LYOPH

TSPAN8/TM4SF3 Antibody (458811) [Unconjugated] Summary

• Immunogen: NS0 mouse myeloma cell line transfected with human TSPAN8
  Met1-Lys237 (Ile35Val)
  Accession # AAH05246
• Specificity: Detects human TSPAN8 in Western blots.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Rat
• Gene: TSPAN8
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunocytochemistry 3-25 ug/mL
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for TSPAN8/TM4SF3 Antibody (458811) [Unconjugated]

• CO-029
• tetraspanin 8
• TM4SF3
• TM4SF3tspan-8
• Transmembrane 4 superfamily member 3tetraspanin-8
• TSPAN8
• Tspan-8
• Tumor-associated antigen CO-029

Background

Tetraspanin-8 (TSPAN8), also known as TM4SF3 and human tumor-associated antigen CO-0029, is a member of the transmembrane 4 superfamily. It is a cell surface 27‑34 kDa glycoprotein with 4 transmembrane segments, and a large extracellular loop (LEL) domain from amino acids (aa) 106‑206. Both the N- and C-termini are intracellular. TSPAN8 is expressed by multiple tumor types as well as smooth and skeletal muscle cells, endothelial cells, hematopoietic progenitor cells and non-keratinized squamous epithelium. As with other TSPAN family molecules, TSPAN8 acts as an organizer of microdomains in membranes. Molecules that may be found in these domains include CD151 (another TSPAN family member), beta 1 Integrins, Integrin alpha 4 beta 6, EpCAM, and CD13. TSPAN8 would appear to remove Integrins from the cell surface, facilitating cell motility which, in the case of tumor cells, results in metastasis. TSPAN8 shares 53% aa sequence identity with both mouse and rat TSPAN8.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for TSPAN8/TM4SF3 Antibody (MAB4734)(1)

Publication using MAB4734

• K Dissanayak, M Nõmm, F Lättekivi, Y Ressaissi, K Godakumara, A Lavrits, G Midekessa, J Viil, R Bæk, MM Jørgensen, S Bhattachar, A Andronowsk, A Salumets, Ü Jaakma, A Fazeli Individually cultured bovine embryos produce extracellular vesicles that have the potential to be used as non-invasive embryo quality markers Theriogenology, 2020;149(0):104-116. 2020 [PMID: 32259747] (Microarray, Mouse)

Bioinformatics

• Gene Symbol: TSPAN8
• Uniprot:
  • AAH05246()