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Anti p ampkα thr172

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Anti-p-AMPKα(Thr172) is a lab equipment product produced by Cell Signaling Technology. It is an antibody that detects phosphorylation of AMP-activated protein kinase alpha (AMPKα) at threonine 172.

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Lab products found in correlation

Anti ampkα, by Cell Signaling Technology (9 mentions) Anti β actin, by Cell Signaling Technology (2 mentions) Anti vps34, by Cell Signaling Technology (2 mentions) Anti p raptor ser792, by Cell Signaling Technology (2 mentions) Anti flag m2 hrp, by Merck Group (2 mentions) Anti beclin 1, by Santa Cruz Biotechnology (2 mentions) Anti myc hrp sc40 hrp, by Santa Cruz Biotechnology (2 mentions) Anti uvrag, by Cell Signaling Technology (2 mentions) Anti atg14, by Cell Signaling Technology (2 mentions) Anti tbc1d1, by Cell Signaling Technology (2 mentions) Anti p acc ser79, by Merck Group (2 mentions) Anti acc, by Merck Group (2 mentions) Anti p tbc1d1 ser237, by Merck Group (2 mentions) Anti ha high affinity hrp, by Roche (2 mentions) Anti β actin hrp, by Santa Cruz Biotechnology (2 mentions) Anti raptor, by Cell Signaling Technology (2 mentions) Anti atg14, by MBL Life Science (2 mentions) Mini protean tgx precast gel, by Bio-Rad (2 mentions) Pvdf membrane, by Merck Group (2 mentions) Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

AMPKα (280 citations) P-AMPKα (127 citations) Anti-AMPKα (120 citations) P-AMPKα (Thr172) (66 citations) Anti-p-AMPKα (25 citations)

10 protocols using anti p ampkα thr172

1

Western Blot Analysis of Hepatic Proteins

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Hepatic total lysates were separated by SDS-PAGE and transferred on to a nitrocellulose membrane in an electrophoretic transfer cell (Bio-Rad, USA). At room temperature, the membranes were washed by TBST and blocked with 5% skimmed milk, then incubated with the following antibodies: anti-p-AMPKα-Thr172, anti-AMPKα, anti-SREBP-1c, anti-PPARα, anti-β-actin (Cell Signaling, Danvers, MA, USA) overnight at 4°C. After incubated with horseradish peroxidase-conjugated secondary antibody (Boster Bio-Engineering Co., Ltd), the membranes were washed and detected using diaminobenzidine (DAB) reagent (Tiangen, China).
Fang J., Wang F., Song H., Wang Z., Zuo Z., Cui H., Jia Y., Deng J., Yu S., Hu Y., Shen L., Ma X., Ren Z, & Gou L. (2018). AMPKα pathway involved in hepatic triglyceride metabolism disorder in diet-induced obesity mice following Escherichia coli Infection. Aging (Albany NY), 10(11), 3161-3172.
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2

Antibodies for Western Blot Analysis

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All primary antibodies used for western blots were purchased from commercial sources. Antibodies from Santa Cruz include: anti-beclin 1 (sc11427), anti-myc HRP (sc40 HRP) and anti-β-actin HRP (sc47778 HRP). Antibodies from Cell Signaling Technology include: anti-UVRAG (#11315), anti-Vps34 (#4263), anti-Atg14 (#96752, to detect mouse Atg14), anti-p-AMPKα(Thr172) (#2535), anti-AMPKα (#2532), anti-p-Raptor(Ser792) (#2083), anti-Raptor (#2280), anti-TBC1D1 (#66433) and anti-HA (#3724, to detect Tlr9-HA in tissue lysates of Tlr9-HA KI mice). Other primary antibodies include: anti-p-ACC(Ser79) (#07–303, Millipore), anti-ACC (#04–322, Millipore), anti-p-TBC1D1(Ser237) (#07–2268, Millipore), anti-Atg14 (M184–3, MBL International, to detect human ATG14), anti-HA high affinity-HRP (#12013819001, Roche, to detect overexpressed TLR9-HA in cells and Tlr9-HA after immunoprecipitation from muscle lysates of the Tlr9-HA KI mice), anti-Flag M2-HRP (A8592, Sigma), and anti-GLUT4 (GT41-A, Alpha Diagnonstic International). To detect beclin 1 in Tlr9-HA immunoprecipitates from muscle lysates by western blot analysis, 10% MINI-PROTEAN TGX Precast gels (Bio-Rad) were used. For detection of all other proteins, 4–20% Precast gels were used.
Liu Y., Nguyen P.T., Wang X., Zhao Y., Meacham C.E., Zou Z., Bordieanu B., Johanns M., Vertommen D., Wijshake T., May H., Xiao G., Shoji-Kawata S., Rider M.H., Morrison S.J., Mishra P, & Levine B. (2020). Tlr9 and Beclin 1 Cross-Talk Regulates Muscle AMPK Activation in Exercise. Nature, 578(7796), 605-609.
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3

Western Blot Analysis of AMPK and MAPK Signaling

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Liver tissue was homogenized in HEPES buffer and proteins were separated by SDS-PAGE and transferred to PVDF membranes. Then, membranes were immunoblotted over night at 4°C using the following antibodies: anti-p-AMPKα (Thr172, #2535), anti-AMPKα (#2603), anti-p-SAPK/JNK (Thr183/Tyr185), anti-p-p38 MAP kinase (Thr180/Tyr182, #9211), and anti-p-p44/42 MAPK (Erk1/2, Thr202/Tyr204, #9101); the above antibodies were all purchased from Cell Signaling (Danvers, MA); anti-eNOS (610296) was purshased form Transduction Laboratories (Lexington KY) and anti-b-actin (A5316) was purshased from Sigma Chemical (St. Louis, MO, USA). After washing, bound antibody was detected after incubation for 1 h at room temperature with the corresponding secondary antibody linked to horseradish peroxidase. Bound complexes were detected and quantified by scanning densitometry.
Bejaoui M., Pantazi E., Folch-Puy E., Panisello A., Calvo M., Pasut G., Rimola A., Navasa M., Adam R, & Roselló-Catafau J. (2015). Protective Effect of Intravenous High Molecular Weight Polyethylene Glycol on Fatty Liver Preservation. BioMed Research International, 2015, 794287.
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4

Western Blot Analysis of Zebrafish Liver and HEK293T Cells

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The zebrafish livers at fasting 9 dpf and cultured HEK293T cells were lysed using RIPA lysis. For Western blot as previously described [54 (link),55 (link)]. The protein samples was resolved on a 10% or 12% SDS-PAGE gel, then transferred onto a PVDF membrane (Millipore). Western blot was performed using anti-pRS6 (S240/244) (1:1000; #2215 Cell Signaling, USA), anti-RS6 (1:1000; #2217 Cell Signaling, USA), anti-LC3B (1:1000; ab192890, Abcam), anti-pAKT(Ser473) (1:500; #4046 Cell Signaling, USA), anti-AKT (1:1000; #4691 Cell Signaling, USA), anti-FTCD (1:200; #93327 Cell Signaling, USA), anti-pAMPKα(Thr172) (1:500; #2535 Cell Signaling, USA), anti-AMPKα (1:1000; #2532 Cell Signaling, USA), anti-α-tubulin (1:2000, Santa Cruz, USA), anti-mouse-HRP (1:2000, Abcam, UK) and anti-rabbit-HRP (1:2000, Abcam, UK).
Zhang W., Wu C., Ni R., Yang Q., Luo L, & He J. (2021). Formimidoyltransferase cyclodeaminase prevents the starvation-induced liver hepatomegaly and dysfunction through downregulating mTORC1. PLoS Genetics, 17(12), e1009980.
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5

AMPK and GPR120 Signaling Pathway

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Anti-tGFP (TA150039) was purchased from OriGene (Rockville, MD). Anti-AMPKα (#5831), anti-pAMPKα-Thr172 (#2535), Anti-AMPKα1 (#2795), Anti-AMPKα2 (#2757), anti-c-myc (#5605), anti-pc-myc-Ser62 (#13748), anti-UBC9 (#4786), anti-IL-6 (#13797), anti-NaK ATPase (#7074), anti-α-smooth muscle actin (SMA; #56856), anti-SUMO2/3 (#4971), and anti-MCP-1 (#12838) were purchased from Cell Signaling Technology Inc. (Danvers, MA). Anti-β-arrestin 2 (C16D9) rabbit mAb (Cell Signaling Technology) for the western blotting analysis, and sc-365445 (Santa Cruz Biotechnology) for the IP assay. Anti-β-arrestin 1 (D8O3J) rabbit mAb (Cell Signaling Technology). Anti-α-actin (sc-47778) and anti-SUMO1 (sc-5380) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX). Anti-GPR120 (sc-390752) and anti-SUMO2/3 immunoprecipitation (IP) beads (#BK-162) were purchased from Cellskeleton, Inc and antibody (sc-50331) were purchased from Santa Cruz Biotechnology, Inc. GW9508 (G9797) and AICAR (A9978) were purchased from Sigma-Aldrich (St. Louis, MO). DHA (CAS6217-54-5) was purchased from Cayman Chemical Company (Ann Arbor, MI). Anti-CD68 (14-0681-80) was purchased from Affymetrix-ebioscience.
Yan C.H., Liu H.W., Tian X.X., Li J., Ding Y., Li Y., Mei Z., Zou M.H, & Han Y.L. (2022). AMPKα2 controls the anti-atherosclerotic effects of fish oils by modulating the SUMOylation of GPR120. Nature Communications, 13, 7721.
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6

Western Blot Analysis of Protein Expression

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The whole proteins from jejunums of mice and FHs74Int cells were extracted with RIPA lysis buffer (Beyotime Biotechnology). BCA protein assay kit (Beyotime Biotechnology) was used to determine the concentration of protein. Proteins were separated with SDS-PAGE and transferred to PVDF membranes (Merck Millipore, Darmstadt, Germany) followed by blocking with 5% skim milk (BD/Difco, Sparks, MD, USA). The various primary antibodies used in the experiments were as follows: anti-GPX4 (1:1000; Protein Tech Group, Wuhan, China), anti-ACSL4 (1:1000; ABclonal, Wuhan, China), anti-pSTAT1 (Tyr701) (1:1000; Cell Signaling Technology, Beverly, MA, USA), anti-STAT1 (1:1000; Protein Tech Group), anti-IRF1 (1:1000; ABclonal), anti-AMPKα (1:1000; Cell Signaling Technology), anti-pAMPKα (Thr172) (1:1000; Cell Signaling Technology) and anti-β-actin (1:1000; Protein Tech Group). After incubation with primary antibodies and washing with TBST, membranes were incubated with IRDye-conjugated secondary antibodies (1:10000; Li-COR Biosciences, Lincoln, NE, USA). Images of immunoreactive bands were captured with Odyssey CLx Infrared Imaging system (Li-COR Biosciences) and quantified with ImageJ software (National Institutes of Health).
Kong P., Yang M., Wang Y., Yu K.N., Wu L, & Han W. (2023). Ferroptosis triggered by STAT1- IRF1-ACSL4 pathway was involved in radiation-induced intestinal injury. Redox Biology, 66, 102857.
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7

Antibodies for Western Blot Analysis

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All primary antibodies used for western blots were purchased from commercial sources. Antibodies from Santa Cruz include: anti-beclin 1 (sc11427), anti-myc HRP (sc40 HRP) and anti-β-actin HRP (sc47778 HRP). Antibodies from Cell Signaling Technology include: anti-UVRAG (#11315), anti-Vps34 (#4263), anti-Atg14 (#96752, to detect mouse Atg14), anti-p-AMPKα(Thr172) (#2535), anti-AMPKα (#2532), anti-p-Raptor(Ser792) (#2083), anti-Raptor (#2280), anti-TBC1D1 (#66433) and anti-HA (#3724, to detect Tlr9-HA in tissue lysates of Tlr9-HA KI mice). Other primary antibodies include: anti-p-ACC(Ser79) (#07–303, Millipore), anti-ACC (#04–322, Millipore), anti-p-TBC1D1(Ser237) (#07–2268, Millipore), anti-Atg14 (M184–3, MBL International, to detect human ATG14), anti-HA high affinity-HRP (#12013819001, Roche, to detect overexpressed TLR9-HA in cells and Tlr9-HA after immunoprecipitation from muscle lysates of the Tlr9-HA KI mice), anti-Flag M2-HRP (A8592, Sigma), and anti-GLUT4 (GT41-A, Alpha Diagnonstic International). To detect beclin 1 in Tlr9-HA immunoprecipitates from muscle lysates by western blot analysis, 10% MINI-PROTEAN TGX Precast gels (Bio-Rad) were used. For detection of all other proteins, 4–20% Precast gels were used.
Liu Y., Nguyen P.T., Wang X., Zhao Y., Meacham C.E., Zou Z., Bordieanu B., Johanns M., Vertommen D., Wijshake T., May H., Xiao G., Shoji-Kawata S., Rider M.H., Morrison S.J., Mishra P, & Levine B. (2020). Tlr9 and Beclin 1 Cross-Talk Regulates Muscle AMPK Activation in Exercise. Nature, 578(7796), 605-609.
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8

Protein Expression Analysis by Western Blot

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Protein was extracted from cells cultured in 6-well plates using RIPA lysis buffer (Thermo Fisher Scientific, 89900) with protease inhibitors (Thermo Fisher Scientific, 78440). A total of 30-μg protein lysates was separated by 4%–12% NuPage Bis-Tris gels (Thermo Fisher Scientific, NP0336) and then transferred to polyvinylidene fluoride membranes (Bio-Rad, 1620264). The membranes were blocked with 5% BSA (Sigma-Aldrich, A7906–100G), probed with primary antibodies overnight at 4°C, and then incubated with the relevant secondary antibody for 1 hour at room temperature. Bands were visualized using ECL (Thermo Fisher Scientific, 34080) with Bio-Rad imaging system. The antibodies of anti-PARP (9532), anti-Rb (9309), anti-pRbSer807/811 (8516), anti-γH2AX (9718), anti-TFAM (8076), anti-AMPKα (5831), anti-pAMPKαThr172 (2535), anti-ERRα (13826), anti-VDAC (4661), anti-PHB1 (2426), anti-HSP60 (12165), anti-NRF2 (12721), anti-SDHA (11998), anti–β-actin (12620), anti–rabbit-IgG-HRP (7074), and anti–mouse-IgG-HRP (7076) were purchased from Cell Signaling Technology. The antibodies of anti-PGC1α (66369), anti-UQCRC1 (21705), anti-ATP5A1 (14676), anti-COXIV (11242), anti-NDUFB8 (14794), and anti-TUFM (26730) were purchased from Proteintech. Two or three independent experiments were performed.
Wei S., Yin D., Yu S., Lin X., Savani M.R., Du K., Ku Y., Wu D., Li S., Liu H., Tian M., Chen Y., Bowie M., Hariharan S., Waitkus M., Keir S.T., Sugarman E.T., Deek R.A., Labrie M., Khasraw M., Lu Y., Mills G.B., Herlyn M., Wu K., Liu L., Wei Z., Flaherty K.T., Abdullah K., Zhang G, & Ashley D.M. (2022). Antitumor Activity of a Mitochondrial-Targeted HSP90 Inhibitor in Gliomas. Clinical cancer research : an official journal of the American Association for Cancer Research, 28(10), 2180-2195.
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9

Western Blotting Procedure for Protein Analysis

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20 μg of cells or 40 μg of tissue lysates were subjected to electrophoresis on 10% acrylamide gels and transferred to PVDF membranes. The membranes were incubated for 1 h with blocking buffer (either TBS-T containing 5% [w/v] BSA or 5% skim milk). The membranes were then incubated with the indicated primary antibodies diluted in blocking buffer (1:1,000) for 12 h at 4 °C: anti-pAmpk-α Thr172 (Cell Signaling, #2535s), anti-Ampk-α1 (R&D, #AF3197), anti-Ampk-α (Cell Signaling, #2532), anti-Ampk-α2 (abcam, ab3760), anti-UCP1 (abcam, ab10983), anti-HSP90 (Cell Signaling, #4874), anti-Bcl2 (abcam, ab196495), anti-cleaved Caspase 3 (Cell Signaling, #9661), anti-GAPDH (Cell Signaling, #2118), anti-Reg3γ (abcam, ab198216), and anti-Reg3α (abcam, ab202057). The membranes were washed three times with TBS-T and incubated with the secondary HRP-conjugated antibodies mouse anti-rabbit IgG-HRP (Cell Signaling, #7074S) (diluted 1:2000 in 5% skim milk) at room temperature for 1 h. Finally, the membranes were washed in TBS-T three times for 10 min each, and the signal was detected using enhanced chemiluminescence reagent (Pierce, IL, USA). Protein levels were quantified by densitometry using Image J software.
Zhang E., Jin L., Wang Y., Tu J., Zheng R., Ding L., Fang Z., Fan M., Al-Abdullah I., Natarajan R., Ma K., Wang Z., Riggs A.D., Shuck S.C., Yang L, & Huang W. (2022). Intestinal AMPK modulation of microbiota mediates crosstalk with brown fat to control thermogenesis. Nature Communications, 13, 1135.
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10

Western Blot Assay for Protein Analysis

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Western blot assay was carried out as described previously [52 (link)]. The following primary antibodies were used at 1:1000 dilutions: rabbit anti-p-eNOS ser1177, anti-eNOS, anti–p-AMPKα thr172, anti-AMPKα, anti-caveolin-1, and anti-β-actin purchased from Cell Signaling Technology (Beverly, MA); Anti-cavin-1 and anti-LDLR purchased from Proteintech (China). The goat anti-rabbit and goat anti-mouse secondary antibodies from Abbkine (Redlands, CA, USA) were used at 1:10 000 dilution.
Bai X.L., Yang X.Y., Li J.Y., Y.e.-.L.i., Jia X., Xiong Z.F., Wang Y.M, & Jin S. (2017). Cavin-1 regulates caveolae-mediated LDL transcytosis: crosstalk in an AMPK/eNOS/ NF-κB/Sp1 loop. Oncotarget, 8(61), 103985-103995.
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