Human Laminin alpha 4 Antibody

AF7340 1 Citation Datasheet / COA / SDS

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AF7340
AF7340-SP

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Human Laminin alpha 4 Antibody Summary

Species Reactivity

Human

Specificity

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.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human Laminin alpha 4
Gln826-Ala1816, predicted
Accession # EAW48267

Formulation

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.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

2 µg/mL

See below

Simple Western

20 µg/mL

See below

Immunohistochemistry

3-25 µg/mL

See below

Immunocytochemistry

5-15 µg/mL

See below

Neutralization

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.

Data Examples

Western Blot

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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.

Immunocytochemistry

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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.

Immunohistochemistry

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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.

Simple Western

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Detection of Human Laminin alpha 4 by Simple Western^TM. 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.

Neutralization

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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 ND₅₀is typically 1.5-7.5 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

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

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.

Entrez Gene IDs

3910 (Human); 16775 (Mouse); 309816 (Rat)

Alternate Names

DKFZp686D23145; LAMA3LAMA4*-1; LAMA4; laminin alpha 4 chain; Laminin alpha 4; laminin subunit alpha-4; laminin, alpha 4; Laminin-14 subunit alpha; Laminin-8 subunit alpha; Laminin-9 subunit alpha

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COA

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Citation for Human Laminin alpha 4 Antibody

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