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Human Laminin alpha 4 Antibody

Catalog #: AF7340
3 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF7340
AF7340-SP
Detection of Human Laminin alpha 4 by Western Blot.
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Citations (3)
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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 either lyophilized or 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.

Scientific Data

Western Blot Detection of Human Laminin a4 antibody by Western Blot. View Larger

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 Laminin a4 antibody in T98G Human Cell Line by Immunocytochemistry (ICC). View Larger

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 Laminin a4 antibody in Human Placenta by Immunohistochemistry (IHC-P). View Larger

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 Detection of Human Laminin a4 antibody by Simple Western<sup>TM</sup>. View Larger

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.

Neutralization Adhesion Induced by Laminin alpha 4 and Neutralization by Human Laminin alpha 4 Antibody. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
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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 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC 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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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.

3 Citations: Showing 1 - 3
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  1. 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
    Theranostics, 2020-08-13;10(22):10274-10289.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  2. Laminin ?4 overexpression in the anterior lens capsule may contribute to the senescence of human lens epithelial cells in age-related cataract
    Authors: Y Yan, H Yu, L Sun, H Liu, C Wang, X Wei, F Song, H Li, H Ge, H Qian, X Li, X Tang, P Liu
    Aging (Albany NY), 2019-05-10;11(9):2699-2723.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry, Migration Assay
  3. 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, 2014-03-27;36(0):5-14.
    Species: Human
    Sample Types: Antibody
    Applications: Neutralization

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