BMX Overexpression Lysate Summary
Description |
BMX Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC202002
Accession#: NM_203281
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 |
BMX |
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 |
77.8 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 BMX Overexpression Lysate
Background
The Tec family of non-receptor tyrosine kinases is composed of six proteins designated Tec, Emt (also known as Itk or Tsk), Btk (previously known as Atk, BPK or Emb), Bmx, Txk (also known as Rlk) and Dsrc28C. All members of the family contain SH3 and SH2 domains and, with the exception of Txk and Dsrc28C, also contain a pleckstrin homology (PH) and a Tec homology (TH) domain in their amino termini. Four alternatively spliced forms of Tec are found to be expressed broadly in cells of hematopoietic lineage and hepatocytes. The 72 kDa Emt gene product associates with CD28 and becomes activated subsequent to CD28 ligation. Btk is necessary for proper B cell development, and mutations in the gene encoding Btk have been associated with families suffering from X-linked agammaglobulinemia, also referred to as Bruton's disease. The 80 kDa Bmx protein shares a high degree of homology with Btk and seems to be expressed at highest levels in the heart. Txk expression is T cell-specific, while expression of the Drosophila Tec homolog, Dsrc28C, is developmentally regulated.
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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