Human Laminin alpha 4 Antibody Summary
Accession # EAW48267
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.
Laminin alpha 4 in Human Placenta. Laminin a4 was detected in immersion fixed paraffin-embedded sections of human placenta using Sheep Anti-Human Laminin a4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7340) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Sheep IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC006). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to Golgi apparatus. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of Human Laminin alpha 4 by Western Blot. Western blot shows lysates of human placenta tissue. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human Laminin a4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7340) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Laminin a4 at approximately 200-220 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Laminin alpha 4 in T98G Human Cell Line. Laminin a4 was detected in immersion fixed T98G human glioblastoma cell line, induced (upper panel) or not (lower panel) with StemXVivo EMT Inducing Media Supplement (Catalog # CCM017), using Sheep Anti-Human Laminin a4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7340) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and cell surfaces. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human Laminin alpha 4 by Simple WesternTM. Simple Western lane view shows lysates of human placenta tissue, loaded at 0.2 mg/mL. A specific band was detected for Laminin a4 at approximately 233 kDa (as indicated) using 20 µg/mL of Sheep Anti-Human Laminin a4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7340) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.
Adhesion Induced by Laminin alpha 4 and Neutralization by Human Laminin alpha 4 Antibody. Recombinant Human Laminin a4 (Catalog # 7340-A4) supports adhesion in the HT1080 human fibrosarcoma cell line in a dose-dependent manner (orange line), as measured by Calcein AM. Adhesion elicited by Recombinant Human Laminin a4 (5 µg/mL) is neutralized (green line) by increasing concentrations of Sheep Anti-Human Laminin a4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7340). The ND50 is typically 1.5-7.5 µg/mL.
Preparation and Storage
- 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: Laminin alpha 4
Laminin alpha 4 (LAMA-4 [Laminin subunit alpha 4]; also Laminin-14/423, -8/411 and -9/421 subunit alpha) is a 180-230 kDa secreted member of the laminin family of molecules. It is found in the basement membranes of adipocytes, endothelial cells, cardiac and visceral smooth muscle cells, fibroblasts, keratinocytes, and pancreatic acinar cells. Laminin is a heterotrimeric glycoprotein that attaches cells to basement membranes. It is composed of covalently-linked alpha -, beta - and gamma -chains, and is known to attach cells to basement membranes. In the case of the alpha -subunit, this is mediated through binding to multiple integrins ( alpha 3 beta 1, alpha v beta 3 and alpha 6 beta 1), plus dystroglycan and the syndecans-2 and -4. Mature human Laminin alpha 4 is 1799 amino acids (aa) in length (aa 25-1823). It contains multiple domains, including four EGF‑like domains (aa 82-255) and five Laminin G-like domains (aa 833-1820). There is a chondroitin sulfate attachment at the N‑terminus that accounts for 20-30 kDa of MW. Laminin alpha 4 is cleaved between the Laminin G-like 3 and 4 domains, creating a soluble 44 kDa fragment that possesses antibacterial activity. There are two potential splice variants, one that shows a deletion of aa 266-272, and another that contains a 55 aa substitution for aa 66-1823. Over aa 826-1816, human Laminin alpha 4 shares 91% aa sequence identity with mouse Laminin alpha 4.
Citations for Human Laminin alpha 4 Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
Citations: Showing 1 - 2
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LAMA4 upregulation is associated with high liver metastasis potential and poor survival outcome of Pancreatic Cancer
Authors: B Zheng, J Qu, K Ohuchida, H Feng, SJF Chong, Z Yan, Y Piao, P Liu, N Sheng, D Eguchi, T Ohtsuka, K Mizumoto, Z Liu, S Pervaiz, P Gong, M Nakamura
Sample Types: Whole Cells
Monoclonal antibodies to human laminin alpha4 chain globular domain inhibit tumor cell adhesion and migration on laminins 411 and 421, and binding of alpha6beta1 integrin and MCAM to alpha4-laminins.
Authors: Ishikawa T, Wondimu Z, Oikawa Y, Ingerpuu S, Virtanen I, Patarroyo M
Matrix Biol, 2014;36(0):5-14.
Sample Types: Antibody
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