ZNF148 Overexpression Lysate Summary
| Description |
ZNF148 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC222687
Accession#: NM_021964
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 |
ZNF148 |
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 |
88.8 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 ZNF148 Overexpression Lysate
Background
ZBP-89, also known as BFCOL1, BERF1 and ZNF 148, is a zinc finger transcription factor that is universally expressed. ZBP-89, a Kruppel-like repressor protein, is the silencer element binding factor for Vimentin. ZBP-89 has been shown to bind to GC-rich DNA elements in promoters for gastrin, ornithine decarboxylase and the cyclin-dependent kinase inhibitor p21 (also designated Cip1 or WAF1). ZBP-89 expression is induced by trans-retinoic acid or butyrate, which also induces terminal differentiation of colon cancer cells. ZBP-89 cooperates with histone acetyltransferase coactivator p300 in the regulation of p21, a cyclin-dependent kinase inhibitor whose associated gene is a target gene of p53. ZBP-89 also regulates cell proliferation, in part, through its ability to directly bind the p53 protein and retard its nuclear export. Elevated levels of ZBP-89 induce growth arrest and apoptosis in human gastrointestinal cells.
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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