TM4SF1/L6 Antibody

Western blot shows lysates of LL/2 mouse Lewis lung carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Mouse TM4SF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7514) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for TM4SF1 at approximately 26 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

TM4SF1 was detected in immersion fixed frozen sections of mouse embryo (13 d.p.c.) using Sheep Anti-Mouse TM4SF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7514) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to the trabeculae of the developing heart. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Laminin promotes the interaction between TM4SF1 and integrin alpha 6 in Eca109 cells.A Con- and TM4SF1 overexpressed- Eca109 cell lysates (under laminin-coating condition) were immunoblotted by anti-p-FAK, anti-FAK, anti-p-PI3K p85 (Try458), anti- PI3K p85, anti-pAKT, and anti-AKT antibodies. B List of TM4SF1-associated cell membrane proteins in Eca109 cells under laminin-coating condition, identified by mass spectrometric analysis. C Co-IP of endogenous TM4SF1 with endogenous integrin alpha 6, beta 4, and beta 1 in cell lysate. TM4SF1 was immunoprecipitated from Eca109 cells, which were cultured on the dishes with or without laminin-coating, and immunoblotted with antibodies against integrin alpha 6, beta 4, beta 1, and TM4SF1. The input was immunoblotted with antibodies against integrin alpha 6, beta 4, beta 1, p-FAK, FAK, TM4SF1, and beta -actin. D Co-IP of endogenous TM4SF1 with endogenous integrin alpha 6, beta 4, and beta 1 in ESCC patient tissues. TM4SF1 was immunoprecipitated from homogenized ESCC patient tissues lysate and immunoblotted with antibodies against integrin alpha 6, beta 4, beta 1, and TM4SF1. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35835740), licensed under a CC-BY license. Not internally tested by R&D Systems.

The Laminin-TM4SF1-integrin alpha 6-FAK signaling axis mediates ESCC cell migration. A–C Immunoblotting of p-FAK, FAK, TM4SF1, integrin alpha 6, and beta -actin in (left panel) and migration ability of (right panel) TM4SF1-overexpressed, integrin alpha 6-overexpressed, and TM4SF1/integrin alpha 6 double overexpressed Eca109 cells (A), Con-, TM4SF1-knockdown-, related TM4SF1 rescued-, integrin alpha 6- overexpressed, and TM4SF1-rescued/integrin alpha 6- overexpressed KYSE-410 cells (B), and integrin alpha 6-knockdown- and related alpha 6 rescued- Con or TM4SF1 overexpressed Eca109 cells (C) under laminin coating condition (LE: long exposure; SE: short exposure). Representative photos were taken, and then the migrated cells were counted. Statistical significance was determined by a two-tailed unpaired t test. Error bars are means ± s.d. (right panel; n = 3; *P < 0.05; **P < 0.01; ***P < 0.001). D Immunoblotting of p-FAK, FAK, TM4SF1, integrin alpha 6, and beta -actin in (upper panel) and migration ability of (lower panel) TM4SF1-overexpressed, integrin alpha 6-overexpressed, and TM4SF1/integrin alpha 6 double overexpressed KYSE-410 cells pre-treated with or without VS-4718 (1 μM) for 24 h. Representative photos were taken and then the migrated cells were counted. Statistical significance was determined by a two-tailed unpaired t test. Error bars are means ± s.d. (right panel; n = 3; n.s., not statistically significant; *P < 0.05; ***P < 0.001). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35835740), licensed under a CC-BY license. Not internally tested by R&D Systems.

Laminin promotes the interaction between TM4SF1 and integrin alpha 6 in Eca109 cells.A Con- and TM4SF1 overexpressed- Eca109 cell lysates (under laminin-coating condition) were immunoblotted by anti-p-FAK, anti-FAK, anti-p-PI3K p85 (Try458), anti- PI3K p85, anti-pAKT, and anti-AKT antibodies. B List of TM4SF1-associated cell membrane proteins in Eca109 cells under laminin-coating condition, identified by mass spectrometric analysis. C Co-IP of endogenous TM4SF1 with endogenous integrin alpha 6, beta 4, and beta 1 in cell lysate. TM4SF1 was immunoprecipitated from Eca109 cells, which were cultured on the dishes with or without laminin-coating, and immunoblotted with antibodies against integrin alpha 6, beta 4, beta 1, and TM4SF1. The input was immunoblotted with antibodies against integrin alpha 6, beta 4, beta 1, p-FAK, FAK, TM4SF1, and beta -actin. D Co-IP of endogenous TM4SF1 with endogenous integrin alpha 6, beta 4, and beta 1 in ESCC patient tissues. TM4SF1 was immunoprecipitated from homogenized ESCC patient tissues lysate and immunoblotted with antibodies against integrin alpha 6, beta 4, beta 1, and TM4SF1. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35835740), licensed under a CC-BY license. Not internally tested by R&D Systems.

The Laminin-TM4SF1-integrin alpha 6-FAK signaling axis mediates ESCC cell migration. A–C Immunoblotting of p-FAK, FAK, TM4SF1, integrin alpha 6, and beta -actin in (left panel) and migration ability of (right panel) TM4SF1-overexpressed, integrin alpha 6-overexpressed, and TM4SF1/integrin alpha 6 double overexpressed Eca109 cells (A), Con-, TM4SF1-knockdown-, related TM4SF1 rescued-, integrin alpha 6- overexpressed, and TM4SF1-rescued/integrin alpha 6- overexpressed KYSE-410 cells (B), and integrin alpha 6-knockdown- and related alpha 6 rescued- Con or TM4SF1 overexpressed Eca109 cells (C) under laminin coating condition (LE: long exposure; SE: short exposure). Representative photos were taken, and then the migrated cells were counted. Statistical significance was determined by a two-tailed unpaired t test. Error bars are means ± s.d. (right panel; n = 3; *P < 0.05; **P < 0.01; ***P < 0.001). D Immunoblotting of p-FAK, FAK, TM4SF1, integrin alpha 6, and beta -actin in (upper panel) and migration ability of (lower panel) TM4SF1-overexpressed, integrin alpha 6-overexpressed, and TM4SF1/integrin alpha 6 double overexpressed KYSE-410 cells pre-treated with or without VS-4718 (1 μM) for 24 h. Representative photos were taken and then the migrated cells were counted. Statistical significance was determined by a two-tailed unpaired t test. Error bars are means ± s.d. (right panel; n = 3; n.s., not statistically significant; *P < 0.05; ***P < 0.001). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35835740), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu
• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

TM4SF1/L6 Antibody Summary

• Immunogen: E. coli-derived recombinant mouse TM4SF1
  Ala116-Thr161
  Accession # Q64302
• Specificity: Detects mouse TM4SF1 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: TM4SF1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 1 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: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

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

Alternate Names for TM4SF1/L6 Antibody

• L6
• M3s1
• M3S1Tumor-associated antigen L6
• Membrane component chromosome 3 surface marker 1
• membrane component, chromosome 3, surface marker 1
• TAAL6
• TAAL6H-L6
• TM4SF1
• transmembrane 4 L six family member 1
• transmembrane 4 L6 family member 1
• transmembrane 4 superfamily member 1

Background

TM4SF1 (Transmembrane 4 L6 family Member 1; also L6 and M3s1) is a 23-28 kDa member of the L6 tetraspanin family of molecules. It is expressed by fibroblasts, endothelial cells and a variety of tumor cells. TM4SF1 is embedded in both the plasma membrane and the membrane of late endocytic organelles. Here, it appears to be ubiquitinated, and to regulate endocytosis, an action that impacts effective cell migration. TM4SF1 is reported to interact with alpha 5 and beta 1 integrins, and this may impact cell motility. It also suppresses the expression of CD63 and CD82, two molecules that impede cell mobility. Mouse TM4SF1 is a 202 amino acid (aa) 4‑transmembrane (TM) glycoprotein. It contains a short N-terminal cytoplasmic region (aa 1-9), followed by four TM regions and another C-terminal cytoplasmic region (aa 183-202). There is one potential isoform variant that shows an alternative start site at Met60. Over aa 116-161, mouse TM4SF1 shares 73% and 96% aa sequence identity with human and rat TM4SF1, respectively.

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 TM4SF1/L6 Antibody (AF7514)(3)

Publications using AF7514

• Liu, M;Vandekeere, A;Liu, XZ;Schenck, S;Fernández-García, J;Tricot, T;Peng-Winkler, Y;Demicco, M;Broekaert, D;Vermeire, I;Theile, J;Zels, G;Pabba, A;Desmedt, C;Brunner, JD;Altea-Manzano, P;Fendt, SM; Palmitoylation-mediated regulation of KAT2A promotes lung metastasis in breast cancer Nature cell biology 2026-03-13 [PMID: 41826695] (Flow Cytometry, Mouse)
• S Hou, X Hao, J Li, S Weng, J Wang, T Zhao, W Li, X Hu, B Deng, J Gu, Q Hang TM4SF1 promotes esophageal squamous cell carcinoma metastasis by interacting with integrin alpha6 Oncogene, 2022-07-14;13(7):609. 2022-07-14 [PMID: 35835740] (Western Blot, Human)
• WJ Zacharias, DB Frank, JA Zepp, MP Morley, FA Alkhaleel, J Kong, S Zhou, E Cantu, EE Morrisey Regeneration of the lung alveolus by an evolutionarily conserved epithelial progenitor Nature, 2018-02-28;555(7695):251-255. 2018-02-28 [PMID: 29489752] (Flow Cytometry, Mouse)

Bioinformatics

• Gene Symbol: TM4SF1
• Uniprot:
  • Q64302()