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BID Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-10076

Novus Biologicals, part of Bio-Techne
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NBP2-10076

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for BID Overexpression Lysate

BID Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC221674

Accession#: NM_197967

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.

Product Specifications

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.

TMW

11.1 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.

Type

Overexpression

Scientific Data Images for BID Overexpression Lysate

Western Blot: BID Overexpression Lysate [NBP2-10076]

Western Blot: BID Overexpression Lysate [NBP2-10076]

Western Blot: Bid Overexpression Lysate (Adult Normal) [NBP2-10076] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Bid.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: BID

The Bcl-2 family of apoptosis-related genes plays central roles in regulating apoptotic pathways (reviewed in Thomadaki and Scorilas, 2006). Regulation of cell death through apoptosis is critical for the maintenance of homeostasis, defense against infectious agents, and normal development. Bcl-2 family proteins regulate apoptosis primarily through the regulation of mitochondrial outer membrane permeability. In mammals, the family consists of both prosurvival (antiapoptotic) and proapoptotic (prodeath) members. Cellular homeostasis is thought to be dependent on a balance between the actions of prosurvival and proapoptotic proteins. Bcl-2 family proteins can be divided into 3 main subfamilies on the basis of their function and the content of their Bcl-2 homology (BH) domains, for example: 1) Prosurvival: Bcl-2, Bcl-XL, Bcl-W, A1, and Mcl-1 2) Proapoptotic (multidomain): Bax, Bak, and Bok. 3) BH3-only (proapoptotic): Bad, Bcl-XS, Bid, Bik, Bim, Blk, Bmf, Bnip, Noxa, and Puma. Prosurvival members inhibit cells from undergoing apoptosis, whereas proapoptotic and BH3-only subfamily members promote apoptosis. There are 4 BH domains (1-4) conserved among Bcl-2 family proteins. The BH domains are important for function as well as for heterodimerization between family members. Typical prosurvival family members have all four BH domains (1-4), whereas proapoptotic (multidomain) members have BH1, 2 and 3 domains and BH3-only members have only the BH3 domain. Overall, the relative ratio of prosurvival and proapoptotic proteins determines the suseptibility of a cell to various apoptotic stimuli. Many Bcl-2 family proteins are differentially expressed in various malignancies and some are useful prognostic biomarkers. Prosurvival proteins are often elevated in diverse cancers and have the potential to confer resistance to both endogenous cell death stimuli and cancer treatments. Alterations in the ratio or levels of Bcl-2 family proteins have been also associated with nonmalignant diseases including neurodegenerative diseases, autoimmune diseases, AIDs, Down's syndrome, cardiovascular diseases, diabetes, glomerulonephritis, and muscular dystrophy. IMG-5693 recognizes Bid (approx. 19-23 kDa) and cleaved/truncated forms of Bid. The C-terminal cleaved/truncated form (11-15 kDa) of Bid is often referred to as tBid in the literature (reviewed in Yin, 2006).

Long Name

BH3 Interacting Domain Death Agonist

Alternate Names

apoptic death agonist, BH3 interacting domain death agonist, BH3-interacting domain death agonist, BID isoform ES(1b), BID isoform L(2), BID isoform Si6, desmocollin type 4, FP497, Human BID coding sequence, MGC15319, MGC42355, p22 BID, tbid

Gene Symbol

BID

Additional BID Products

Product Documents for BID Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for BID Overexpression Lysate

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.

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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