Anti mkp3

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-MKP3 is a purified antibody that recognizes the MKP3 (MAP kinase phosphatase 3) protein. MKP3 is a dual-specificity phosphatase that inactivates extracellular signal-regulated kinases (ERKs).

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Lab products found in correlation

Anti phospho erk1 2, by Cell Signaling Technology (2 mentions) Criterion gel, by Bio-Rad (2 mentions) Odyssey clx imaging system, by LI COR (2 mentions) Anti gfp, by Thermo Fisher Scientific (2 mentions) Anti flag, by Merck Group (2 mentions) Trans blot turbo blotting system, by Bio-Rad (2 mentions) Anti rcan1, by Merck Group (2 mentions) Anti erk1 2, by Cell Signaling Technology (2 mentions) Anti lamin a c, by Cell Signaling Technology (2 mentions) Anti myc, by Cell Signaling Technology (2 mentions) Anti ets2, by Santa Cruz Biotechnology (2 mentions) Anti nfatc1, by Thermo Fisher Scientific (2 mentions) Anti gapdh, by Biosynth (2 mentions) Anti phospho akt, by Cell Signaling Technology (1 mentions) Protease inhibitor and phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions) M per buffer, by Thermo Fisher Scientific (1 mentions) Okadaic acid, by Merck Group (1 mentions) Anti akt, by Cell Signaling Technology (1 mentions) Anti phospho mek1 2, by Cell Signaling Technology (1 mentions)

4 protocols using anti mkp3

Western Blotting Analysis of Signaling Pathways

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Whole‐cell lysates were prepared in M‐PER buffer supplemented with protease inhibitor and phosphatase inhibitor (Thermo Fisher Scientific, Waltham, MA, USA), resolved by SDS/PAGE, and blotted with indicated antibodies. SuperSignal West Pico Chemiluminescent Substrate and SuperSignal Western Blot Enhancer (Thermo Fisher Scientific) were used to enhance blotting signal when needed. The following antibodies were used in this study: anti‐MAP4K4, anti‐phospho‐ERK1/2, anti‐phospho‐JNK, anti‐JNK, anti‐phospho‐p38, anti‐p38, anti‐AKT, anti‐phospho‐AKT, anti‐MEK1/2, anti‐phospho‐MEK1/2, anti‐RAS and anti‐PP2A‐C (Cell Signaling Technology, Danvers, MA, USA); anti‐ERK1/2, anti‐GAPDH, anti‐β‐actin, anti‐PP2A‐B56β, and anti‐MKP3 (Santa Cruz Biotechnology, Dallas, TX, USA); anti‐6Χ HIS (Immunology Consultants Laboratory, Inc., Lake Oswego, OR, USA). Erlotinib and trametinib were purchased from Selleck Chemicals (Houston, TX, USA). The protein phosphatase 2 (PP2A) inhibitor okadaic acid (OA) was purchased from Sigma Aldrich. The PP2A activator FTY720 was purchased from Cayman Chemical (Ann Arbor, MI, USA). MAP4K4 inhibitor PF‐06260933 was purchased from Tocris Bioscience (Avonmouth, Bristol, UK). EGF was obtained from Thermo Fisher Scientific.

Gao X., Chen G., Gao C., Zhang D.H., Kuan S., Stabile L.P., Liu G, & Hu J. (2017). MAP4K4 is a novel MAPK/ERK pathway regulator required for lung adenocarcinoma maintenance. Molecular Oncology, 11(6), 628-639.

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Ras/MAPK Pathway Antibody Panel Analysis

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The primary antibodies used were as follows: anti-H-Ras (sc-29, Santa Cruz Biotech Inc., Santa Cruz, CA, USA), anti-K-Ras (Sc-30, Santa Cruz), anti-N-Ras (sc-31, Santa Cruz), anti-Erk1 (sc-94, Santa Cruz), anti-pErk1/2 (Thr²⁰²/Tyr²⁰⁴, Cat. No. 4370, Cell Signaling Technology, Danvers, MA, USA), anti-(sc-1619, Santa Cruz), anti-pAkt (Ser⁴⁷³, Cat. No. 4060, Cell signaling), anti-MKP3 (sc-377070, Santa Cruz), anti-α-tubulin (sc-398103, Santa Cruz), and anti-lamin B (sc-374015, Santa Cruz). Secondary antibodies were from KPL Inc. (Gaithersburg, MD, USA). Human FGF-basic (hFGF-basic, Cat. No. 8910) and U0126 (Cat. No. 9903) were purchased from Cell Signaling Technology.

Koo J., Wang S., Kang N., Hur S.J, & Bahk Y.Y. (2016). Induction of MAP kinase phosphatase 3 through Erk/MAP kinase activation in three oncogenic Ras (H-, K- and N-Ras)-expressing NIH/3T3 mouse embryonic fibroblast cell lines. BMB Reports, 49(7), 370-375.

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Protein Fractionation and Western Blot Analysis

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Proteins were prepared with radioimmunoprecipitation assay buffer (50 mmol/L Tris-HCl, pH 7.4, 150 mmol/L NaCl, 1 mmol/L EDTA, 0.25% sodium deoxycholate, 0.1% SDS) supplemented with protease and phosphatase inhibitors (A32961; Thermo Fisher). Nuclear and cytoplasmic fractions were isolated using a kit according to the manufacturer’s instructions (78835; Thermo Fisher). A total of 20 to 40 µg of proteins were separated using Criterion gels (Bio-Rad) and transferred to nitrocellulose membranes using the Trans-Blot Turbo Blotting System (Bio-Rad). The membranes were then blocked with 5% nonfat milk or bovine serum albumin, followed by incubation with primary antibodies overnight at 4°C. After 3 washes, the membranes were incubated with secondary antibody and scanned with an Odyssey CLx Imaging system (LI-COR Biosciences). The following antibodies were used: anti-ETS2 (sc-365666; Santa Cruz Biotechnology), anti-phospho-ETS2 (44-1105G; Thermo Fisher), anti-GAPDH (10R-G109a; Fitzgerald), anti-RCAN1.4 (D6694; Sigma), anti-MKP3 (sc-137246; Santa Cruz Biotechnology), anti-lamin A/C (2032; Cell Signaling), anti-NFATc1 (nuclear factor of activated T cells 1; MA3-024; Thermo Fisher), anti-GFP (green fluorescent protein; A-11120; Invitrogen), anti-FLAG (7425; Sigma), anti-Myc (2276; Cell Signaling), anti-Erk1/2 (9107; Cell Signaling), and anti-phospho-Erk1/2 (9101; Cell Signaling).

Luo Y., Jiang N., May H.I., Luo X., Ferdous A., Schiattarella G.G., Chen G., Li Q., Li C., Rothermel B.A., Jiang D., Lavandero S., Gillette T.G, & Hill J.A. (2021). Cooperative Binding of ETS2 and NFAT Links Erk1/2 and Calcineurin Signaling in the Pathogenesis of Cardiac Hypertrophy. Circulation, 144(1), 34-51.

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Western Blot Analysis of Protein Signaling

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Proteins were prepared with radio-immunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl PH 7.4, 150 mM NaCl, 1mM EDTA, 0.25% sodium deoxycholate, 0.1% SDS) supplemented with protease and phosphatase inhibitors (Thermo Fisher, A32961). Nuclear and cytoplasmic fractions were isolated using a kit according to the manufacturer’s instructions (Thermo Fisher, 78835). 20–40 μg of proteins were separated using Criterion gels (Biorad) and transferred to nitrocellulose membranes using the Trans-Blot^® Turbo^™ Blotting System (Biorad). The membranes were then blocked with 5% non-fat milk or BSA, followed by incubation with primary antibodies overnight at 4°C. After 3 washes, the membranes were incubated with secondary antibody, and scanned with an Odyssey CLx Imaging system (LI-COR). The following antibodies were used: anti-ETS2 (Santa Cruz Biotechnology, sc-365666), anti-p-ETS2 (Thermo Fisher, 44–1105G), anti-GAPDH (Fitzgerald, 10R-G109a), anti-Rcan1.4 (Sigma, D6694), anti-MKP3 (Santa Cruz Biotechnology, sc-137246), anti-Lamin A/C (Cell Signaling, 2032), anti-NFATc1(Thermo Fisher, MA3–024), anti-GFP (Invitrogen, A-11120), anti-flag (Sigma, 7425), anti-myc (Cell Signaling, 2276), anti-Erk1/2 (Cell Signaling, 9107), anti-p-Erk1/2 (Cell Signaling, 9101).

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