Livin Overexpression Lysate

Name: Livin Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-06455

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-06455

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Livin Overexpression Lysate

Livin Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC215363

Accession#: NM_022161

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

30.7 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.

Type

Overexpression

Scientific Data Images for Livin Overexpression Lysate

Western Blot: Livin Overexpression Lysate [NBP2-06455]

Western Blot: Livin Overexpression Lysate (Adult Normal) [NBP2-06455] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Livin.

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: Livin

Livin (BIRC7/KIAP) is a member of the family of inhibitor of apoptosis proteins (IAP) and contains a single copy of a baculovirus IAP repeat (BIR) as well as a RING-type zinc finger domain (reviewed in Ma et al 2006 and Crnkovic-Mertens et al 2006). Resistance towards apoptosis is a hallmark of cancer cells, and overexpression of IAPs can contribute to the development of cancer though inhibiting apoptosis (reviewed in Wright et al. 2005). IAP proteins inhibit apoptosis by binding to and inhibiting caspases through their BIR domain(s). Some IAP members, like Liven, also have a RING-type finger motif at their carboxyl-terminal which may enhance anti-apoptotic activity. Many RING finger-containing IAPs possess E3 ubiquitin ligase activity, and it has been shown that Liven acts an E3 ubiquitin ligase for targeting the degradation of Smac/DIABLO. Smac/DIABLO functions by inhibiting IAP-caspase interactions, thereby promoting apoptosis. Thus degradation of Smac/DIABLO is thought to be a mechanism to enhance Livin anti-apoptotic activity, thereby promoting cell survival. Two splicing variants of Livin, alpha and beta, have been identified. The two isoforms are thought to have different anti-apoptotic properties; however, there are conflicting reports are to whether they actually differ in their biological activities. There is accumulating evidence that Liven plays a significant role in a spectrum of tumor types and it is thought that Liven may have potential as a diagnostic and prognostic tumor marker. Liven is expressed at low levels in adult tissues, and relatively higher levels in developmental tissues and in many cancer cells. Certain cancer patients develop autoantibodies against liven and in a number of cancers, Liven is detected only in the tumors but not, or to substantially lower levels, in the corresponding normal adjacent tissue. This antibody recognizes Livin, both isoform alpha and isoform beta. Human Livin isoform alpha is a 298 amino acid (aa) protein, GenBank no. NP_647478. Human Livin isoform beta is a 280 aa protien, GenBank no. NP_071444.1. The antibody will also recognize other isoforms of Livin that contain the peptide immunogen sequence.

Alternate Names

BIRC7, KIAP, ML-IAP

Gene Symbol

BIRC7

Additional Livin Products

Product Documents for Livin Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for Livin 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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