Noxa Overexpression Lysate

Name: Noxa Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-14534

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-14534

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Noxa Overexpression Lysate

Noxa Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC202071

Accession#: NM_021127

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.

Specificity

This Noxa antibody was developed by immunizing mice with a fusion protein containing human Noxa.

TMW

5.8 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.

Type

Overexpression

Scientific Data Images for Noxa Overexpression Lysate

Western Blot: Noxa Overexpression Lysate [NBL1-14534]

Western Blot: Noxa Overexpression Lysate [NBL1-14534] - (Adult Normal) [NBL1-14534] Left lane: Empty vector transfected control cell lysate (HEK293); Right lane: Over-expression lysate for Noxa.

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: Noxa

Noxa, phorbol-12-myristate-13-acetate-induced protein 1, PMA-induced protein 1, PMAIP1 (Human 6 kDa and Mouse 11.6 kDa) is a Bcl2 homology 3 (BH3) domain containing protein which plays a role in the regulation of apoptosis, specifically a pro-apoptotic role. Human Noxa contains only one BH3 domain, while the mouse and rat proteins contain two BH3 domains. The human protein is encoded by Exon 1 and 3, with Exon 2 present only in two splice variants. Human Noxa splice variants lack a BH3 domain, have no known function, and are rapidly targeted for degradation (1).

Noxa mediated apoptosis may follow its transcriptional activation by p53 as part of the DNA damage response. However, Noxa expression may be induced by HIF-1 alpha under hypoxia, promoting apoptosis independently of p53 (1). Inhibition of the anti-apoptotic Bcl2 family member Myeloid cell leukemia-1 (Mcl-1) by Noxa, leads to the activation of pro-apoptotic Bcl2 homologous antagonist killer (Bak) and Bcl2 associated X (Bax) proteins, and apoptosis through the intrinsic mitochondrial pathway (2). Other BH3 only proteins such as Bim, Puma, Bad and Bid act via the same mechanism, targeting Mcl-1 and inducing Bak/Bax mediated mitochondrial outer membrane permeabilization (MOMP), cytochrome c release and caspase 9 activation (3). Overexpression of antiapoptotic Bcl2 family members is common in various types of cancer including prostate cancer. A recent study identified antiapoptotic Bcl2 proteins involved in the development of resistance towards the androgen receptor antagonist enzalutamide, which is used for the treatment of metastatic castration-resistant prostate cancer (mCRPC) (3).

References

1.Ploner, C., Kofler, R., & Villunger, A. (2008). Noxa: At the tip of the balance between life and death. Oncogene. https://doi.org/10.1038/onc.2009.46

2.Xiang, W., Yang, C. Y., & Bai, L. (2018). MCL-1 inhibition in cancer treatment. OncoTargets and Therapy. https://doi.org/10.2147/OTT.S146228

3.Pilling, A. B., & Hwang, C. (2019). Targeting prosurvival BCL2 signaling through Akt blockade sensitizes castration-resistant prostate cancer cells to enzalutamide. Prostate. https://doi.org/10.1002/pros.23843

Alternate Names

adult T cell leukemia-derived PMA-responsive, APR, Immediate-early-response protein APR, NOXAPMA-induced protein 1, phorbol-12-myristate-13-acetate-induced protein 1, PMA-Induced Protein 1, Protein Noxa

Gene Symbol

PMAIP1

Additional Noxa Products

Product Documents for Noxa Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for Noxa 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.

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