Caspase-9 Overexpression Lysate

Novus Biologicals | Catalog # NBP2-05935

Novus Biologicals
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Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Caspase-9 Overexpression Lysate

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.

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.

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.

Type

Overexpression

Scientific Data Images for Caspase-9 Overexpression Lysate

Western Blot: Caspase-9 Overexpression Lysate [NBP2-05935]

Western Blot: Caspase-9 Overexpression Lysate [NBP2-05935]

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.

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: Caspase-9

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.

Alternate Names

CASP9, Caspase9

Gene Symbol

CASP9

Additional Caspase-9 Products

Product Documents for Caspase-9 Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for Caspase-9 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.

Related Research Areas

Cysteine Proteases and Regulators

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FAQs for Caspase-9 Overexpression Lysate

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    • Q: Could I use HeLa cell lysate as a positive control for Caspase 9 primary antibodies in WB?
      A: 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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