Bcl rambo Overexpression Lysate Summary
Description |
Bcl rambo Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC200042
Accession#: NM_015367
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 |
BCL2L13 |
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 |
52.5 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 Bcl rambo Overexpression Lysate
Background
Apoptosis plays a major role in normal organism development, tissue homeostasis, and removal of damaged cells. Disruption of this process has been implicated in a variety of diseases such as cancer. Members of the Bcl-2 family are known to be critical regulators of this process. These proteins are characterized by the presence of several conserved motifs termed Bcl-2 homology (BH) domains. A novel, widely expressed member termed Bcl-rambo was recently identified. This protein is localized to mitochondria in mammalian cells and its overexpression induces apoptosis which could be blocked by co-expression of inhibitor of apoptosis proteins (IAPs) such as XIAP, cIAP1, and cIAP2. Bcl-rambo shows overall homology to the anti-apoptotic members containing BH motifs, but unlike Bcl-2, the C-terminal membrane anchor of Bcl-rambo is preceded by a unique 250 amino acid insertion. This region by itself can induce apoptosis more efficiently than the Bcl-2 homology regions, suggesting that Bcl-rambo may be important other pro-apoptotic pathways.
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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