Detects human Laminin alpha 4 in direct ELISAs and Western blots. In direct ELISAs, approximately 15% cross-reactivity with recombinant mouse Laminin alpha 4 is observed.
Polyclonal Sheep IgG
Chinese hamster ovary cell line CHO-derived recombinant human Laminin alpha 4 Gln826-Ala1816, predicted Accession # EAW48267
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
<0.10 EU per 1 μg of the antibody by the LAL method.
Measured by its ability to neutralize Laminin alpha 4-induced adhesion in the HT1080 human fibrosarcoma cell line. The Neutralization Dose (ND50) is typically 1.5-7.5 µg/mL in the presence of 5 µg/mL Recombinant Human Laminin alpha 4.
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.
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 alpha 4 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 alpha 4 at approximately 200‑220 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Laminin alpha 4 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 alpha 4 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.
Adhesion Induced by Laminin alpha 4 and Neutralization by Human Laminin alpha 4 Antibody. Recombinant Human Laminin alpha 4 (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 alpha 4 (5 µg/mL) is neutralized (green line) by increasing concentrations of Sheep Anti-Human Laminin alpha 4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7340). The ND50 is typically 1.5-7.5 µg/mL.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
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
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: 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.
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