Snail Overexpression Lysate Summary
Description |
Snail Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC204581
Accession#: NM_005985
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT. |
Gene |
SNAI1 |
Applications/Dilutions
Dilutions |
|
Application Notes |
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag ( NBP1-71705) present on the protein construct. Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading. |
Theoretical MW |
28.9 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
Packaging, Storage & Formulations
Storage |
Store at -80C. Avoid freeze-thaw cycles. |
Buffer |
RIPA buffer |
Lysate Details for Array
Notes
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
Alternate Names for Snail Overexpression Lysate
Background
Snail, also called SNAIL1 or SNAI1, is a zinc-finger transcription factor belonging to the Snail superfamily and encoded by the SNAI1 gene (1,2). Snail was first discovered in Drosophila and has homologs in many species including vertebrates and humans (1,2). The Snail family members includes Snail (Snail1), Slug (Snail2), and Smuc (Snail3) (1,2). In humans, Snail is expressed in a number of tissues including placenta, brain, and skeletal muscle, but is most highly expressed by the kidneys (1). Snail functions in repression of E-cadherin transcription which is associated with epithelial-mesenchymal transition (EMT) that is especially prominent during embryonic development (1-5). Along with Snail, other related EMT-inducing transcription factors (EMT-TFs) include the Twist and ZEB protein families (3). Snail is synthesized as a protein of 264 amino acids (aa) with an N-terminal SNAG domain, a serine-rich domain (SRD), nuclear export sequences (NES), and four C-terminal zinc-finger binding domains, with a theoretical molecular weight of 29 kDa (1,3). Snail activity is largely regulated through post-translational modifications such as phosphorylation, ubiquitination, and glycosylation, which impacts Snail's localization and stability, amongst other things (1-3, 5).
In addition to its role in embryonic development, Snail-induced EMT is also associated with cancer metastasis (1-5). Snail is expressed in a variety of cancer lines including breast cancer, cervical carcinoma, and colorectal carcinoma, and typically results in increased migration, invasion, and metastasis (1). Accordingly, Snail expression is also correlated with drug resistance and tumor recurrence (1-5). Chemical inhibitors that target Snail have shown some promise in reducing or eliminating Snail-induced EMT, increasing E-cadherin expression, and increasing tumor regression (1).
1. Kaufhold, S., & Bonavida, B. (2014). Central role of Snail1 in the regulation of EMT and resistance in cancer: a target for therapeutic intervention. Journal of Experimental & Clinical Cancer Research. https://doi.org/10.1186/s13046-014-0062-0
2. Wang, Y., Shi, J., Chai, K., Ying, X., & Zhou, B. P. (2013). The Role of Snail in EMT and Tumorigenesis. Current Cancer Drug Targets. https://doi.org/10.2174/15680096113136660102
3. Kang, E., Seo, J., Yoon, H., & Cho, S. (2021). The Post-Translational Regulation of Epithelial-Mesenchymal Transition-Inducing Transcription Factors in Cancer Metastasis. International Journal of Molecular Sciences. https://doi.org/10.3390/ijms22073591
4. Seo, J., Ha, J., Kang, E., & Cho, S. (2021). The role of epithelial-mesenchymal transition-regulating transcription factors in anti-cancer drug resistance. Archives of Pharmacal Research. https://doi.org/10.1007/s12272-021-01321-x
5. Baulida, J., Diaz, V. M., & Herreros, A. G. (2019). Snail1: A Transcriptional Factor Controlled at Multiple Levels. Journal of Clinical Medicine. https://doi.org/10.3390/jcm8060757
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are
guaranteed for 6 months from date of receipt.
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Product General Protocols
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Video Protocols
FAQs for Snail Lysate (NBL1-16265). (Showing 1 - 1 of 1 FAQ).
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There are many kinds of antibodies you supply against SNAIL. Could you give me your recommendation? Which is the best for the IHC-P experiments the species of my samples to be tested is human.
- Our SNAIL antibody with catalog # NBP1-19529 has been successfully validated for IHC-P in paraffin-embedded human lung carcinoma tissue and I would highly recommend you to use the same for your samples too. The working dilutions of this antibody for IHC-P ranges from 1:50 - 1:200 and beside IHC, you can use this antibody for Western Blot and Immunofluorescence also.