Noxa Antibody - BSA Free

Western Blot: Noxa Antibody [NB100-56368] - Noxa expression was stably reduced in HCT116 cells by shRNA. Western blot image submitted by a verified customer review.

Immunohistochemistry-Paraffin: Noxa Antibody [NB100-56368] - IHC analysis of formalin fixed paraffin-embedded (FFPE) human tonsil using Noxa antibody at 1:50 on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10 mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 30 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Whole slide scanning and capturing of representative images was performed using Aperio AT2 (Leica Biosystems). Staining was performed by Histowiz.

Immunocytochemistry/Immunofluorescence: Noxa Antibody [NB100-56368] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-Noxa (NB100-56368) antibody at a 1:200 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Western Blot: Noxa Antibody [NB100-56368] - Western blot analysis of Noxa in A) human, B) mouse, and C) rat thymus tissue using Noxa antibody NB100-56368 at 2 ug/mL.

• Reactivity: Hu, Mu
• Applications: WB, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

Noxa Antibody - BSA Free Summary

• Immunogen: This Noxa antibody was developed by immunizing rabbits with a mixture of synthetic peptides containing amino acids 51-66 and 75-90 of mouse Noxa.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: PMAIP1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:200
  • Immunohistochemistry 1:50 - 1:250
  • Immunohistochemistry-Paraffin 1:50 - 1:250. Use reported in scientific literature (Inta et al. (2006))
  • Western Blot 1 - 2 ug/mL
• Theoretical MW: 11.6 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.
• Control:

  Mouse Thymus Whole Tissue Lysate (Adult Whole Normal)
  Noxa Overexpression Lysate
  Human Thymus Whole Tissue Lysate (Adult Whole Normal)

Reactivity Notes

Immunogen displays the following percentage of sequence identity for non-tested species: amino acids 51-66: Rat (77%); amino acids 75-90: Rat (93%), Human (76%).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for Noxa Antibody - BSA Free

• 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

Background

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

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Noxa Antibody (NB100-56368)(6)

Publications using NB100-56368

• JY Baik, Z Liu, D Jiao, HJ Kwon, J Yan, C Kadigamuwa, M Choe, R Lake, M Kruhlak, M Tandon, Z Cai, S Choksi, ZG Liu ZBP1 not RIPK1 mediates tumor necroptosis in breast cancer Nature Communications, 2021-05-11;12(1):2666. 2021-05-11 [PMID: 33976222]
• Eno CO, Zhao G, Venkatanarayan A et al. Noxa couples lysosomal membrane permeabilization and apoptosis during oxidative stress. Free Radic Biol Med 2013-12-01 [PMID: 23770082] (WB, Human)
• Martin LJ, Liu Z, Pipino J et al. Molecular regulation of DNA damage-induced apoptosis in neurons of cerebral cortex. Cereb Cortex. 2009-06-01 [PMID: 18820287] (WB, Mouse)
  
  Details:
  WB (mouse cortical neuron cultures), Figs. 3A, 6A.
• Fei Q, McCormack AL, Di Monte DA, Ethell DW. Paraquat neurotoxicity is mediated by a Bak-dependent mechanism. J Biol Chem. 2008-02-08 [PMID: 18056701] (WB, Mouse)
  
  Details:
  WB (mouse brain tissue) Figs. 3, 4H; Table 1.
• Inta I, Paxian S, Maegele I et al. Bim and Noxa are candidates to mediate the deleterious effect of the NF-kappa B subunit RelA in cerebral ischemia. J Neurosci. 2006-12-13 [PMID: 17167080] (IHC-P)
  
  Details:
  1. Noxa (IMG-451) [IHC (paraffin), Fig.2 (mice brain)].
• McNeill-Blue C, Wetmore BA, Sanchez JF et al. Apoptosis mediated by p53 in rat neural AF5 cells following treatment with hydrogen peroxide and staurosporine. Brain Res. 2006-09-27 [PMID: 16901471] (WB)

Review for Noxa Antibody (NB100-56368) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 04/04/2014

Summary

• Application: Western Blot
• Sample Tested: whole cell lysate of human colon cancer cell HCT116
• Species: Human
• Lot: 072402A-03

Bioinformatics

• Gene Symbol: PMAIP1
• Entrez:
  • 5366(Human)
  • 58801(Mouse)
  • 492821(Rat)
• Uniprot:
  • Q9JM54(Mouse)