Hrk Overexpression Lysate Summary
Description |
Hrk Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC212069
Accession#: NM_003806
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 |
HRK |
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 |
9.7 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 Hrk Overexpression Lysate
Background
Apoptosis or programmed cell death is a physiological cellular process characterized by cell shrinkage, membrane blebbing, DNA fragmentation, and release of Cytochrome C from the mitochondria. It is utilized by the organism to get rid of unwanted cells, which is critical for normal development and homeostasis of an organism. Disregulation of normal apoptosis process have been implicated in a variety of diseases, including cancer, autoimmune diseases, viral infections, etc. Programmed cell death occurs through complex cascades of cell signaling in which Bcl-2 family members, among others, play an important role.The Bcl-2 family of proteins regulate apoptosis as well as execute death signals at the mitochondrion. Members of this family include both pro- and anti-apoptotic proteins that hare homology sequences called Bcl-2 Homology domains (BH1-4) which mediate dimmer formation. The BH3 proteins, such as BID, NOXA, PUMA, BIK, BIM and BAD are all pro-apoptotic and share sequence homology within the amphipathic alpha-helical BH3 region, which is required for their apoptotic function. They may trigger release of death-inducing molecules such as Cytochrome C, Smac, and endonuclease G. Anti-apoptotic family members, including Bcl-2 and Bcl-XL, play inhibitory roles. Bcl-2 family proteins may form homodimers or heterodimers between pro- and anti-apoptotic members, the ratios of which determine the cell fate.
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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