Human Laminin  alpha 4 Antibody

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.

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 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.

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.

Detection of Human Laminin alpha 4 by Western Blot Flow induces remodeling of the endothelial extracellular matrix.A, Proteome and (B) cell surface proteome data of identified laminins. T or CT represents the enzyme used for protein digestion. The depicted numbers represent the biotin-modified lysine residue. C, Schematic representation of laminin 411 with the linker region between domains LG3 and LG4. The blue lysine residues were found to be decreased under flow conditions (as depicted in panel B). D, Immunoblot analysis of LAMA4 in ECs exposed to flow over time. alpha -tubulin was used as a loading control. E, Protein distribution of LAMA5 and LAMA4 after flow-exposure. Green represents LAMA4 or LAMA5, blue is HOECHST. The scale bar is 10 μm. Depicted are maximum intensity projections. F, Proteome and (G) cell surface proteome data of identified fibulins. H, Immunofluorescence analysis of EFEMP1 and FBLN2 abundance and distribution affected by flow. Red is VE-cadherin, blue is HOECHST. Depicted are maximum intensity projections. Micrographs are representative for 3 independent experiments. The scale bar is 10 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32332107), licensed under a CC-BY license. Not internally tested by R&D Systems.