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PUMA Overexpression Lysate

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Western Blot: PUMA Overexpression Lysate (Adult Normal) [NBL1-07925] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PUMA.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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PUMA Overexpression Lysate Summary

Description

PUMA Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC213203

Accession#: NM_014417

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
BBC3

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 (cat# 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
20.4 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 PUMA Overexpression Lysate

  • BCL2 binding component 3
  • bcl-2-binding component 3
  • JFY1
  • JFY-1
  • p53 up-regulated modulator of apoptosis
  • PUMAFLJ42994

Background

p53 upregulated modulator of apoptosis (PUMA), also called BBC3, is a target for activation by the p53 tumor-suppressor gene. p53 functions as a transcriptional activator, and influences p53-inducible genes that play a role in the induction of apoptosis in response to p53. Antisense inhibition of PUMA expression reduced the apoptotic response to p53, and PUMA is likely to play a role in mediating p53-induced cell death through the cytochrome c/Apaf-1-dependent pathway.

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 PUMA Lysate (NBL1-07925) (0)

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Product General Protocols

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Video Protocols

WB Video Protocol

FAQs for PUMA Lysate (NBL1-07925) (0)

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Research Areas for PUMA Lysate (NBL1-07925)

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Blogs on PUMA.

A link between Autophagy and Apoptosis: Chat with First Author Brent E. Fitzwalter
By Christina Towers, PhD. Autophagy is a cellular recycling process and most often a pro-survival mechanism that regulates cellular homeostasis. On the contrary, apoptosis is an extensively conserved and elaborate pr...  Read full blog post.

Pathway Highlight: Which caspase substrates contribute to the morphological features associated with apoptosis?
Apoptosis, or programmed cell death, is controlled by a caspase signal cascade that activates downstream signals to induce the morphological changes used to differentiate apoptosis from other forms of cell death.  Novus Biologicals offers a variet...  Read full blog post.

Altered expression of BCL2 in cancer
Similar to other cell processes, the balance between cell survival and cell death is an important equilibrium that when altered expression of genes can lead to a variety of disease.  For example, too little cell death can promote cell overgrowth a...  Read full blog post.

IRE1 alpha dependent apoptotic-signaling pathway
Despite in depth characterization of the role of IRE1 alpha (inositol-requiring enzyme 1 alpha) in activating the unfolded protein response (UPR) in the ER - little is known about the molecular mechanisms by which this ER protein has shown to reg...  Read full blog post.

Understanding Noxa Regulation of Apoptosis
Noxa is a pro-apoptotic gene belonging to the Bcl2 protein family that is unique in that it contains only BH3 domain. The BH3-only subclass of proteins, including proteins like PUMA and Bim in addition to Noxa, regulate the remaining Bcl-2 family memb...  Read full blog post.

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Bioinformatics

Gene Symbol BBC3