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Caspase-9 Overexpression Lysate

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Western Blot: Caspase 9 Overexpression Lysate (Adult Normal) [NBP2-05935] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Caspase 9.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Caspase-9 Overexpression Lysate Summary

Description

Caspase-9 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC213375

Accession#: NM_032996

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
CASP9

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
30 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 Caspase-9 Overexpression Lysate

  • APAF-3APAF3
  • apoptotic protease activating factor 3
  • Apoptotic protease Mch-6
  • Apoptotic protease-activating factor 3
  • CASP9
  • CASP-9
  • caspase 9, apoptosis-related cysteine peptidase
  • Caspase9
  • Caspase-9
  • EC 3.4.22.62
  • ICE-LAP6CASPASE-9c
  • ICE-like apoptotic protease 6
  • MCH6apoptosis-related cysteine protease

Background

Caspase 9 (CASP9) is a member of the cysteine-aspartic acid protease (caspase) family that is involved in the execution-phase of cell apoptosis. Caspase 9 is expressed as an inactive proenzyme which then undergos proteolytic processing by upstream caspases, to form an active (cleaved) caspase 9 form.

There are two classes of caspases involved in apoptosis; initiators (activation by receptor cluster) and effectors (activation by mitochondrial permeability transition). Caspase 3 is considered an initiator, as initiated caspase-9 goes on to cleave the effector procaspase-3 and procaspase-7.

Caspase 9 antibodies are important tools for apoptosis studies and research on the caspase family mediated apoptosis signaling 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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Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for Caspase-9 Lysate (NBP2-05935). (Showing 1 - 1 of 1 FAQ).

  1. Could I use HeLa cell lysate as a positive control for Caspase 9 primary antibodies in WB?
    • After looking through our testing it does appear you should be fine using HeLa as a positive control cell lysate for Caspase 9 in Western Blot, as we have used this sample successfully in our QC tests and shown expression of Caspase 9.

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Research Areas for Caspase-9 Lysate (NBP2-05935)

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By Rachel M.A. Linger, Ph.D.What is survivin?Survivin is a small (16.5 kDa) protein normally found in human fetal tissue. In contrast, survivin is typically undetectable in most normal adult tissues. Expression of ...  Read full blog post.

Caspase-3, The Executioner of Apoptosis
The Role of Caspase-3 in ApoptosisCaspase-3 enzyme is a member of the family of endoproteases which regulate inflammation and apoptosis signaling networks. Caspase-3 is known as an executioner caspase in apoptosis because of its role in coordinat...  Read full blog post.

Caspase 9 - an important apoptosis marker
Caspases are essential mediators of programmed cell death and are needed for both the induction of apoptosis as well as for aiding the degradation of cellular structures. Initiator caspases (such as Caspase-9) sense and respond to various signals i...  Read full blog post.

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Bioinformatics

Gene Symbol CASP9