FOXL1 Overexpression Lysate Summary
Description |
FOXL1 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC216442
Accession#: NM_005250
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 |
FOXL1 |
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 |
36.3 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 FOXL1 Overexpression Lysate
Background
FOXL1 is a 337 amino acid protein encoded by the mouse gene Foxl1. FOXL1 belongs to the fork head family and contains one fork head DNA-binding domain. The HNF3/fork head family includes a large number of transcription factors that share a structurally related DNA binding domain. Fork head factors are known to play important roles both during development and in adults. FOXL1 is a winged helix transcriptional regulator expressed in the mesenchymal layer of developing and mature gastrointestinal tract. FOXL1- deficient mice exhibit various defects not only in the epithelial layer of the gastrointestinal tract but also in gut-associated lymphoid tissues. In the small intestine of FOXL1-deficient mice, the formation of Peyer's patches is affected, particularly in the caudal region. FOXL1 is a mesenchymal modifier of the adenomatous polyposis coli (APC) gene products and plays a key role in gastrointestinal tumorigenesis.
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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