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BID Overexpression Lysate

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Western Blot: Bid Overexpression Lysate (Adult Normal) [NBL1-07979] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Bid.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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BID Overexpression Lysate Summary

Description

BID Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC207261

Accession#: NM_001196

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
BID

Applications/Dilutions

Dilutions
  • Western Blot
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
21.8 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

Type
Overexpression

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 BID Overexpression Lysate

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

Background

This gene encodes a death agonist that heterodimerizes with either agonist BAX or antagonist BCL2. The encoded protein is a member of the BCL-2 family of cell death regulators. It is a mediator of mitochondrial damage induced by caspase-8 (CASP8); CASP8 cleaves this encoded protein, and the COOH-terminal part translocates to mitochondria where it triggers cytochrome c release. Multiple alternatively spliced transcript variants have been found, but the full-length nature of some variants has not been defined. [provided by RefSeq]. Transcript Variant: This variant (2) has an alternate 5' exon and uses a downstream in-frame start codon, as compared to variant 1. The encoded isoform 2 thus has a shorter N-terminus, as compared to isoform 1. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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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Publications for BID Lysate (NBL1-07979) (0)

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Product General Protocols

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Video Protocols

WB Video Protocol

FAQs for BID Lysate (NBL1-07979) (0)

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Research Areas for BID Lysate (NBL1-07979)

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Apoptosis and Necroptosis Part I: Important factors to identify both types of programmed cell death
Different types of cell death have classically been identified by discrete morphological changes. The hallmarks of apoptosis include cell shrinkage, nuclear fragmentation and membrane blebbing whereas necroptosis is characterized by cell swelling ...  Read full blog post.

The use of apoptosis antibodies and controls in cell death research
Apoptosis is a method of programmed cell death that is notably characterized by a morphological change in cellular nuclei and membrane appearance.  Not to be confused with necrosis, apoptosis is a pathway that is induced by a variety of factors tha...  Read full blog post.

The role of p53 in UV radiation DNA damage and subsequent tumorogenesis
p53, the protein product of the tp53 gene, is one of the most widely studied tumor suppressor proteins in cancer research.  p53 is unique in that it demonstrates both tumor suppressive and tumor progressive properties depending on whether it is fu...  Read full blog post.

active/cleaved Caspase 2 - Inducing apoptosis in response to cellular stress
Caspase-2 is a highly conserved member of the caspase family involved in the initiation and execution of apoptosis. While its function is still poorly understood, caspase-2 is thought to be important for apoptosis in response to DNA damage, bacteri...  Read full blog post.

Bcl-2 - an antiapoptotic protein with an important role in cancer cell survival
B-cell lymphoma 2 (Bcl-2) protein is an oncogene that normally acts as an apoptotic inhibitor and localizes to the mitochondrial membrane where it prevents the release of cytochrome c. The Bcl-2 protein family consists of over 20 proteins each co...  Read full blog post.

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Bioinformatics

Gene Symbol BID