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Bag-1 Overexpression Lysate

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Western Blot: Bag-1 Overexpression Lysate [NBL1-07901] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Bag1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

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Bag-1 Overexpression Lysate Summary

Description

Bag-1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC201272

Accession#: NM_004323

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
BAG1

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
38.7 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

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 Bag-1 Overexpression Lysate

  • BAG family molecular chaperone regulator 1
  • Bag1
  • Bag-1
  • Bcl-2 associating athanogene-1 protein
  • Bcl-2-associated athanogene 1
  • BCL2-associated athanogene
  • Bcl-2-binding protein
  • glucocortoid receptor-associated protein RAP46,46-KD
  • HAP
  • RAP46

Background

The oncogene BCL2 is a membrane protein that blocks a step in a pathway leading to apoptosis or programmed cell death. The protein encoded by this gene binds to BCL2 and is referred to as BCL2-associated athanogene. It enhances the anti-apoptotic effects of BCL2 and represents a link between growth factor receptors and anti-apoptotic mechanisms. At least three protein isoforms are encoded by this mRNA through the use of a non-AUG (CUG) start site, and alternative, downstream, AUG translation initiation sites. [provided by RefSeq]. 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 Bag-1 Lysate (NBL1-07901) (0)

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Product General Protocols

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Blogs on Bag-1.

BAG3 - Hsp70 is my friend!
The BAG proteins are a large family of chaperone regulators governing a wide range of cell processes such as proliferation, survival, stress response, tumorigenesis, neuronal differentiation, growth arrest and apoptosis as reviewed in Takayama et al1....  Read full blog post.

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Bioinformatics

Gene Symbol BAG1