PKN3 Antibody - BSA Free

Western Blot: PKN3 Antibody [NBP1-30102] - Whole cell lysate from HeLa (5, 15 and 50 ug) and 293T (T; 50 ug) cells. Antibody used at 0.1 ug/ml.

Western Blot: PKN3 Antibody [NBP1-30102] - Image submitted by a verified customer review.

Western Blot: PKN3 Antibody [NBP1-30102] - PKN3 regulates growth in 2D and in 3D environment of Src-transformed MEFs through interaction with p130CaS. Immunoblot of SC cells and SC cells with PKN3 gene inactivated using CRISPR/CAS9 (SCpkn3-/-). Inactivation of PKN3 expression is visualized by antibody anti-PKN3. Antibody anti-actin was used as loading control. Image collected and cropped by CiteAb from the following publication (//onlinelibrary.wiley.com/doi/abs/10.1002/1878-0261.12401), licensed under a CC-BY license.

Western Blot: PKN3 Antibody [NBP1-30102] - Influence of TP Agonist on the Activation of PRK1, PRK2 & PRK3. Panel C. In order to identify the species of PRK subject to U46619-induced T-loop phosphorylation, PC-3 cells initially transfected for 72 hr with 30 nM siRNAPRK1, siRNAPRK2, siRNAPRK3 or, as controls, with a scrambled siRNAControl. Thereafter, cells serum starved & stimulated with U46619 for 60 min or with vehicle & then immunoblotted (20 μg/lane) with anti-phospho-PRK1Thr774/PRK2Thr816/PRK3Thr718 & with anti-PRK1, anti-PRK2, anti-PRK3 or, as loading controls, HDJ2 antisera, as indicated. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.4664), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PKN3 Antibody [NBP1-30102] - Effect of siRNA-mediated disruption of PRK1, PRK2 & PRK3 expression on TP agonist-induced proliferation & migration of PC-3 cellsPanels A-D. PC-3 cells were transfected for 48 hr with 30 nM siRNAPRK1, siRNAPRK2, siRNAPRK3 or, as controls, with a scrambled siRNA (siRNAControl) prior to stimulation with U46619 (10 nM) or vehicle (0.0001% EtOH) for the indicated time specific to the assay, where non-transfected cells served as a reference. Panel A: For analysis of proliferation, 72 hr post-siRNA transfection PC-3 cells were stimulated for 48 hr with U46619 (10 nM) or vehicle (0.0001% EtOH). Panel B: For analysis of colony formation, 48 hr post-siRNA transfection PC-3 cells were stimulated with U46619 (10 nM) or vehicle (0.0001% EtOH) in soft agar & assessed 10 day post-treatment for colony formation. Panel C: For analysis of migration, 72 hr post-siRNA transfection PC-3 cells were stimulated for 8 hr with U46619 (10 nM) or vehicle (0.0001% EtOH). In Panels A-C, the bar charts show mean relative levels of PC-3 cell proliferation, colony formation & migration (± SEM, n ≥ 3), where levels in the vehicle-treated cells are assigned a value of 100%. The asterisks indicate where U46619-stimulation resulted in significant increases in proliferation, colony formation or migration by PC-3 cells compared to vehicle-treated cells, where ** & *** indicates p 50 measurements – 3 per cell; ***P < 0.001, one‐way ANOVA on ranks followed by Dunn's post hoc test). (C) Src‐transformed p130Cas−/−MEFs co‐expressing p130Cas (SC) & mouse Flag tagged PKN3 WT or Flag‐PKN3 mPR are shown. Cells were grown on FN‐coated coverslips for 48 h, fixed, & stained for p130Cas by anti‐pTyr165 p130Cas antibody (pY165 p130Cas; 2nd 405), for actin by Phalloidin 488 & for Flag‐PKN3 by anti‐Flag antibody (2nd 633). Reflection (670 nm) indicates fibronectin degradation. All scale bars represent 20 μm. Cell were imaged by Leica TCS SP8 microscope system equipped with Leica 63×/1.45 oil objective. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30422386), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PKN3 Antibody [NBP1-30102] - PKN3 overexpression regulates growth of MEFs, & this effect requires PKN3–p130Cas interaction. (A) Immunoblotted lysates from MEFs p130Cas−/− or MEFs p130Cas−/− re‐expressing p130Cas (p130Cas+) treated by Doxycycline (Dox) to induce expression of mCherry‐PKN3 or mCherry alone. p130Cas presence detected by ani‐p130Cas antibody & mCherry epitope by anti‐mCherry antibody. (B) Dynamics of mCherry‐PKN3 expression after supplementation w/ Dox shown by immunoblot w/ anti‐mCherry antibody. (C–E) Effect of induced mCherry‐PKN3 expression on cell growth. Representative graphs showing growth of MEFs p130Cas−/− re‐expressing p130Cas (p130Cas+) (C) or MEFs p130Cas−/− (D) measured in real‐time using xCELLigence RTCA (real‐time cell analysis) system instrument. (E) Quantification of cell growth change induced by mCherry‐PKN3 expression (‘−’ indicates inducible mCherry expression used as negative control). Slope ratios reflecting cell growth calculated from log growth phase of cell growth (indicated by dotted lines; see C & D). (F) Immunoblotted lysates from MEFs p130Cas−/− re‐expressing p130Cas (p130Cas+) treated or not treated by Dox which induced expression of Flag‐fused PKN3 variants (WT, mPR, KD, empty vector). Stimulated overexpression of PKN3 detected by anti‐PKN3 antibody & its activity by antibody anti‐pT849 PKN3. (G) Quantification of cell growth change stimulated by Dox‐inducible expression of Flag‐fused PKN3 variants (WT, mPR, KD) in MEFs p130Cas−/− re‐expressing p130Cas (p130Cas+). All error bars indicate means ± SD calculated from 3 to 5 independent experiments (each in triplicates). Statistical significance always calculated between induced & noninduced cells & evaluated by one‐way repeated ANOVA followed by Turkey post hoc test (***P < 0.001). Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/30422386), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu
• Applications: WB, IP (-), KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

PKN3 Antibody - BSA Free Summary

• Description: Novus Biologicals Knockout (KO) Validated Rabbit PKN3 Antibody - BSA Free (NBP1-30102) is a polyclonal antibody validated for use in WB. Anti-PKN3 Antibody: Cited in 6 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: maps to a region between residue 225 and 275 of human protein kinase N3 using the numbering given in entry NP_037487.2
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: PKN3
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Knockout Validated Reactivity validation reported in (PMID: 30518210)
  • Western Blot 1:2000-1:10000
• Theoretical MW: 99 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.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 30422386).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: Tris-Citrate/Phosphate (pH 7.0 - 8.0)
• Preservative: 0.09% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for PKN3 Antibody - BSA Free

• EC 2.7.11
• EC 2.7.11.13
• PKNBETA
• protein kinase N3
• Protein kinase PKN-beta
• Protein-kinase C-related kinase 3
• RP11-545E17.1
• serine/threonine-protein kinase N3

Background

Contributes to invasiveness in malignant prostate cancer

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 PKN3 Antibody (NBP1-30102)(6)

Publications using NBP1-30102

• Arang N, Lubrano S, Ceribelli M et al. High-throughput chemogenetic drug screening reveals PKC-RhoA/PKN as a targetable signaling vulnerability in GNAQ-driven uveal melanoma Cell reports. Medicine 2023-10-13 [PMID: 37858338] (WB, Human)
• Browne CM, Jiang B, Ficarro SB et al. A Chemoproteomic Strategy for Direct and Proteome-Wide Covalent Inhibitor Target-Site Identification J. Am. Chem. Soc. 2019-01-09 [PMID: 30518210] (KO, WB, Human)
• Gemperle J, Dibus M, Koudelkova L et al. The interaction of p130Cas with PKN3 promotes malignant growth Mol Oncol. [PMID: 30422386] (WB, Mouse)
• Aine G O'Sullivan, Eamon P Mulvaney, B Therese Kinsella Regulation of protein kinase C-related kinase (PRK) signalling by the TP alpha and TP beta isoforms of the human thromboxane A 2 receptor: Implications for thromboxane- and androgen- dependent neoplastic and epigenetic responses in prostate cancer. Biochimica et biophysica acta. Molecular basis of disease 2018-11-19 [PMID: 28108419]
• O'Sullivan AG, Mulvaney EP, Hyland PB, Kinsella BT. Protein kinase C-related kinase 1 and 2 play an essential role in thromboxane-mediated neoplastic responses in prostate cancer. Oncotarget 2015-09-22 [PMID: 26296974]
• Collazos A, Michael N, Whelan RD et al. Site recognition and substrate screens for PKN family proteins. Biochem J;438(3):535-43. 2011-09-15 [PMID: 21749319]

Reviews for PKN3 Antibody (NBP1-30102) (2)

Average Rating: 4.5

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Human.

reviewed by: Verified Customer WB Human 06/05/2017

Summary

• Application: Western Blot
• Sample Tested: A375m2 cell lysate,Immunoprecipitation
• Species: Human
• Lot: 1

reviewed by: Verified Customer WB Human 08/05/2014

Summary

• Application: Western Blot
• Species: Human

FAQs for PKN3 Antibody (NBP1-30102). (Showing 1 - 2 of 2 FAQs).

1. A customer needs an antibody to PKN3 which is specific for this isoform and not cross reacting with the forms PKN1 and PKN2. Would this be suitable for NBP1-30102?
   • I ran an alignment of the immunogen sequence for NBP1-30102 against the sequences for PKN1 and PKN2 and the similarity is low. We haven't tested for cross reactivity, but the customer should feel confident that within these regions there is not many amino acids shared between PKN3 and PKN1 and PKN2.
2. What I'm looking for is an antibody that recognizes preferably the N-terminal part of the human and mouse PKN3 kinase (since my protein of interest has been shown to bind to a region just upstream of the kinase domain - between aa 510 and 522). My intention is to use this antibody for Western blotting and IPs. I have found a PKN3 antibody (NBP1-30102) raised against an internal region of the human protein, that can recognize endogenous levels of the protein in HeLa lysates according to your datasheet (Western blot). This antibody however is not recommended for IPs. Does this mean that IPs have been tested and the antibody doesn't work for this application? Since I want to be able to (by Western blotting) detect mouse PKN3, do you know if this antibody is suitable for this?
   • NBP1-30102 recognizes a region from amino acids 225-275 which would be upstream of the kinase domain. Our antibodies are guaranteed for use in all species and applications listed, so for this antibody, it has been shown to work in Western Blots but looks to be unsuitable for IPs. The IP (-) means that we have tested for its use in IP and have been unsuccessful so we do not recommend if for IP. The region recognized shows only 75% sequence similarity in mouse so it may or may not cross-react with mouse but it is something we cannot guarantee. As far as an alternative antibody, our other PKN3 antibodies we currently sell are raised against full-length PKN3 and are not epitope mapped. One of these could be useful for IP and NBP1-30102 could be used to detect.

Bioinformatics

• Gene Symbol: PKN3
• Entrez:
  • 29941(Human)
• Uniprot:
  • Q6P5Z2(Human)