|Detection of Human Snail by Western Blot. Western blot shows lysates of A549 human lung carcinoma cell line and JEG‑3 human epithelial choriocarcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human Snail Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3639) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Snail at approximately 29 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Chromatin Immunoprecipitation (ChIP)
|Detection of Snail-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for 30 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Snail/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human Snail Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3639) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The E-Cadherin promoter was detected by standard PCR.|
|Snail in A549 Human Cell Line. Snail was detected in immersion fixed A549 human lung carcinoma cell line treated with Recombinant Human TGF-beta 1 (left panel, Catalog # 240-B) or untreated (right panel) using Goat Anti-Human Snail Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3639) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
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Immunocytochemistry/Immunofluorescence: Human Snail Antibody [AF3639]
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