Anti pim2

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-Pim2 is a laboratory reagent used to detect the presence and measure the levels of the Pim2 protein in biological samples. Pim2 is a serine/threonine protein kinase that plays a role in cell signaling pathways. The Anti-Pim2 product can be used in various experimental techniques, such as western blotting, immunoprecipitation, and immunohistochemistry, to study the expression and localization of Pim2 in cells or tissues.

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Lab products found in correlation

Anti pim3, by Cell Signaling Technology (5 mentions) Anti pim1, by Cell Signaling Technology (4 mentions) Anti β actin, by Merck Group (2 mentions) Anti akt, by Cell Signaling Technology (2 mentions) Anti phospho eif4b, by Cell Signaling Technology (2 mentions) Azd1208, by AstraZeneca (2 mentions) Anti flag, by Merck Group (2 mentions) Anti 4e bp1, by Cell Signaling Technology (2 mentions) Anti β tubulin, by Merck Group (2 mentions) Hrp linked mouse igg, by GE Healthcare (1 mentions) Anti ha, by Cell Signaling Technology (1 mentions) Hrp conjugated anti β actin, by Merck Group (1 mentions) Azd1208, by Adooq (1 mentions) Pim447, by Novartis (1 mentions) Human recombinant insulin, by Thermo Fisher Scientific (1 mentions) Cycloheximide (chx), by Merck Group (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) Anti ha, by Merck Group (1 mentions) Anti rps6, by Santa Cruz Biotechnology (1 mentions) Anti phospho tyrosine p tyr 100, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Anti-PIM2 (4730), (2 citations)

5 protocols using anti pim2

Probing Protein Kinase Signaling in Stem Cells

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Primary antibodies used for western blotting included anti‐PIM1 (Cell Signaling Technology, Cat # 2907), anti‐PIM2 (Cell Signaling Technology, Cat # 4730), anti‐PIM3 (Cell Signaling Technology, Cat # 4165), anti‐pIRS1‐S1101 (Cell Signaling Technology, Cat # S1101 Cat # 2385), anti‐insulin receptor substrate 1 (IRS1; Cell Signaling Technology, Cat # 06‐248), anti‐HA (Cell Signaling Technology, Cat # 14031), anti‐SOX2 [(E‐4) sc‐365823; Santa Cruz Biotech, Santa Cruz, CA, USA], anti‐OCT‐4 [(C‐10) sc‐5279; Santa Cruz Biotech], and anti‐NANOG [(5A10) sc‐134218; Santa Cruz Biotech]. HRP‐conjugated anti‐β‐actin (Cat # A3854) was purchased from Sigma‐Aldrich. HRP‐linked mouse IgG (Cat # NA931V) and rabbit IgG (Cat # NAV934V) were purchased from GE Healthcare Life Sciences (Princeton, NJ, USA).
Doxycycline (Dox) hydrochloride (Cat # D3447) was purchased from Sigma‐Aldrich. AZD1208 (Cat # A13203) was purchased from Adooq Bioscience (Irvine, CA, USA). Enza (MDV3100; Cat # S1250) was purchased from Selleckchem (Houston, TX, USA). PIM447 was a gift from Novartis (Basel, Switzerland).

Singh N., Padi S.K., Bearss J.J., Pandey R., Okumura K., Beltran H., Song J.H., Kraft A.S, & Olive V. (2020). PIM protein kinases regulate the level of the long noncoding RNA H19 to control stem cell gene transcription and modulate tumor growth. Molecular Oncology, 14(5), 974-990.

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Antibody Utilization in Signaling Pathway Analysis

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The following antibodies were purchased from Cell Signaling Technology: anti-phospho-tyrosine (p-Tyr-100), anti-Pim1, anti-Pim2, anti-Pim3, anti-Cyclin D3, anti-phospho-eIF4B, anti-AKT, anti-phospho-AKT (S473), anti-phospho-AKT (T308), anti-p70S6K, anti-phospho-p70S6K (T389), anti-phospho-S6 (S240/244), anti-phospho-GSK3β (S9), and anti-c-Myc, and anti-phospho IRS1 (S1101, S318, S302, S307, S636/639, S612) antibodies which recognize the following residues in human (h) or mouse (m) IRS1; anti-phospho IRS1 hS1101 (m1097); mS318 (hS323); mS302 (hS307); mS307 (hS312); hS636/S639 (mS632/635); mS612 (hS616). This numbering refers to human IRS1 (P35568); and mouse IRS1 (P35569). Anti-IRS1 (R & D systems), anti-RPS6 (Santa Cruz Biotechnology), anti-His (GenScript), anti-HA and anti-β-actin (Sigma) antibodies were used in these studies. Horseradish peroxidase (HRP)-linked enhanced chemiluminescence (ECL), mouse IgG and rabbit IgG secondary antibodies were purchased from GE Healthcare Life Sciences. Various protein kinase inhibitors, GNE-652 (Genentech), LGB321 (Novartis), AZD1208, AZD5363 (AstraZeneca) and PP242 (Selleck Chemicals) were used in these studies. All other chemicals including cycloheximide (CHX) and PMA were purchased from Sigma. Human recombinant insulin and insulin-like growth factor 1 (IGF1) were obtained from Peprotech.

Song J.H., Padi S.K., Luevano L.A., Minden M.D., DeAngelo D.J., Hardiman G., Ball L.E., Warfel N.A, & Kraft A.S. (2016). Insulin receptor substrate 1 is a substrate of the Pim protein kinases. Oncotarget, 7(15), 20152-20165.

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Antibody and Inhibitor Reagents for Signaling Studies

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The following antibodies were purchased from Cell Signaling Technology: Anti-Pim-1 (Cat#3247), anti-Pim2 (Cat#4730), anti-Pim3 (Cat#4165) anti-MET (Cat#8198), anti-phospho-MET (Cat#3077), anti-BAD (Cat#9239), anti-phospho-BAD (Cat#5284), anti-eIF4B (Cat#3592), anti-phospho-eIF4B (S406, Cat#8151), anti-eIF4G (Cat#2498), anti-eIF4E(Cat#2067), anti-Myc-Tag(Cat#2272), anti-AKT (pan, Cat#4691), anti-phospho-AKT (S473, Cat#4058), anti-phospho-4E-BP1 (Cat#2855), anti-4E-BP1 (Cat#9452), anti-phospho-S6 (Cat#2215), anti-ERK (Cat#9102), anti-phospho-ERK (Cat#9101), anti-Bcl-2(Cat#4223), anti-Bim(Cat#2933), and anti-cleaved PARP(Cat#5625). Anti-β-actin (Cat#A3854), anti-β-tubulin (Cat#T4026), anti-FLAG (Cat#F1804) antibodies were purchased from Sigma. Anti-Mcl-1 antibody (Cat#sc-12756) was from Santa Cruz Biotechnology. A neutralizing antibody against MET was from Abcam (Cat#ab10728). HRP-linked enhanced chemiluminescence (ECL) mouse (Cat#NA931V) and rabbit IgG (Cat#NAV934V) were purchased from GE Healthcare Life Sciences.
The small molecule inhibitors PP242, BEZ235, ABT199, and ABT737 were obtained from Selleck Biochemicals. PHA665752 was from Cayman Chemical. LGB321 was provided by Norvatis. AZD6094 and AZD1208 were provided by AstraZeneca. LY2801653 was from Eli Lilly.

An N., Xiong Y., LaRue A.C., Kraft A.S, & Cen B. (2015). Activation of Pim Kinases Is Sufficient to Promote Resistance to MET Small Molecule Inhibitors. Cancer research, 75(24), 5318-5328.

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Western Blot Protein Analysis Methodology

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Cells were lysed in 2x LSB and heated at 95 °C for 5 min. Proteins were separated by SDS-PAGE, immobilized onto PVDF-membrane (EDM Millipore, Merck) and incubated overnight with anti-PIM-1 (1:500, 12H8; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-PIM-2 (1:1000, D1D2; Cell Signaling Technology, Danvers, MA, USA), anti-PIM-3 (1:1000, D17C9; Cell Signaling Technology), anti-NFATC1 (1:500, Santa Cruz Biotechnology), anti-V5 (1:500, Invitrogen, Carlsbad, CA, USA), anti-Flag (1:500, F1804; Sigma-Aldrich), anti-ACTB (anti-β-actin; 1:1000, 13E5, #4970S, Cell signaling Technology), anti-GAPDH (1:50000, Sigma-Aldrich), anti-β Tubulin (1:40000, Sigma-Aldrich) or anti-Fibrillarin (1:1000, Cell Signaling Technology) antibodies. After incubations with secondary antibodies, chemiluminescence reactions were generated using either Amersham™ ECL Plus or ECL Prime reagents (GE Healthcare).

Eerola S.K., Santio N.M., Rinne S., Kouvonen P., Corthals G.L., Scaravilli M., Scala G., Serra A., Greco D., Ruusuvuori P., Latonen L., Rainio E.M., Visakorpi T, & Koskinen P.J. (2019). Phosphorylation of NFATC1 at PIM1 target sites is essential for its ability to promote prostate cancer cell migration and invasion. Cell Communication and Signaling : CCS, 17, 148.

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Evaluation of Protein Expression Levels

Cited 1 time

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Anti-PIM-1 (Cat# 3247) or Anti-PIM-1 (Cat#2907), anti-PIM-2 (Cat# 4730), anti-PIM-3 (Cat# 4165), anti-c-Myc (Cat# 9402), anti-p21 (Cat# 2947), anti-mTOR (Cat# 2983) and anti-phospho-mTOR (Ser2448) (Cat# 2971), anti-pAKT (Ser473) (Cat# 4058), anti-4EBP1 (Cat# 9644), anti-phospho-S6 (Ser240/244) (Cat# 2215), anti-phospho-4EBP1 (Thr37/46) (Cat# 2855), anti-phospho-eIF4B (Ser406) (Cat# 5399) and anti-αβ-tubulin (Cat #2148) rabbit primary antibodies were purchased from Cell Signaling Technology and diluted as per manufacturer’s protocol. Separation of 50 µg or 30 µg of protein was done with a Western blot and carried out as previously described [12 (link)].

Moore G., Lightner C., Elbai S., Brady L., Nicholson S., Ryan R., O’Sullivan K.E., O’Byrne K.J., Blanco-Aparicio C., Cuffe S., O’Neill M., Heavey S., Finn S.P, & Gately K. (2021). Co-Targeting PIM Kinase and PI3K/mTOR in NSCLC. Cancers, 13(9), 2139.

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