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.
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.
Detection of Snail in A549 Human Cell Line by Flow Cytometry.
A549 human lung carcinoma cell line was stained with Goat Anti-Human Snail Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3639, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Fluorescein-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0109). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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.
Snail is predicted 29 kDa nuclear zinc finger transcriptional repressor that contains an N-terminal basic SNAG domain followed by three classical and one atypical zinc finger domains. During development, Snail is required for the establishment of left-right axis asymmetry. It also regulates the transcription of E-cadherin and other genes involved in epithelial-mesenchymal transitions during cancer progression. Human Snail shares 88% amino acid sequence identity with mouse and rat Snail.
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We have 2 review tested in 1 species: Human.
We have 2 review tested in 1 application: Immunocytochemistry/Immunofluorescence.