- MA5-15815 - Invitrogen Antibodies PRKAA1 antibody

Submitted references ACSL4 contributes to sevoflurane-induced ferroptotic neuronal death in SH-SY5Y cells via the 5' AMP-activated protein kinase/mammalian target of rapamycin pathway.

Cheng L, Zhu X, Liu Y, Zhu K, Lin K, Li F
Annals of translational medicine 2021 Sep;9(18):1454

Antifungal itraconazole ameliorates experimental autoimmune encephalomyelitis through a novel mechanism of action.

Huang H, Tian X, Peng X, Huang L, Mei L, Zhan Y, Chen S, Wu H, Wei G, Cai X
Advances in clinical and experimental medicine : official organ Wroclaw Medical University 2020 May;29(5):535-545

α1AMP-Activated Protein Kinase Protects against Lipopolysaccharide-Induced Endothelial Barrier Disruption via Junctional Reinforcement and Activation of the p38 MAPK/HSP27 Pathway.

Angé M, Castanares-Zapatero D, De Poortere J, Dufeys C, Courtoy GE, Bouzin C, Quarck R, Bertrand L, Beauloye C, Horman S
International journal of molecular sciences 2020 Aug 4;21(15)

Rotating magnetic field delays human umbilical vein endothelial cell aging and prolongs the lifespan of Caenorhabditis elegans.

Xu J, Liu K, Chen T, Zhan T, Ouyang Z, Wang Y, Liu W, Zhang X, Sun Y, Xu G, Wang X
Aging 2019 Nov 22;11(22):10385-10408

Phosphorylation of ULK1 by AMPK regulates translocation of ULK1 to mitochondria and mitophagy.

Tian W, Li W, Chen Y, Yan Z, Huang X, Zhuang H, Zhong W, Chen Y, Wu W, Lin C, Chen H, Hou X, Zhang L, Sui S, Zhao B, Hu Z, Li L, Feng D
FEBS letters 2015 Jul 8;589(15):1847-54

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of AMPK alpha-1 was achieved by transfecting MCF7 cells with AMPK alpha-1 specific validated siRNAs (Silencer® select Product # s100, s101 ). Western blot analysis (Fig. a) was performed using whole cell extracts from the AMPK alpha-1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with AMPK alpha-1 Monoclonal Antibody (Product # MA5-15815, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to AMPK alpha-1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of MCF7 (Lane 1), MDA-MB-231 (Lane 2), LNCaP (Lane 3), PC-3 (Lane 4), NIH/3T3 (Lane 5), HEK-293 (Lane 6), Hep G2 (Lane 7) and A-431 (Lane 8 ). The blot was probed with Anti-AMPK alpha-1 Monoclonal Antibody (Product # MA5-15815, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 63 kDa band corresponding to AMPK alpha-1 was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of AMPK-alpha-1 was performed by loading whole-cell lysates of mouse tissue samples. AMPK-aplha-1 was detected at approximately 62kDa by using a AMPK-alpha-1 monoclonal Antibody (Product # MA5-15815) at a dilution o f1:1000 in Blocking Buffer overnight at 4°C on a rocking platform, followed by a 1-hour room-temperature incubation of a donkey anti-mouse IgG antibody at a dilution of 1:5000. Data courtesy of Thermo Scientific KOL Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of NTERA-2 cells using PRKAA1 monoclonal antibody (Product # MA5-15815) (Green). Blue: DRAQ5 fluorescent DNA dye. Red: actin filaments have been labeled with phalloidin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of AMPK alpha-1 was performed using 70% confluent log phase LNCap cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with AMPK alpha-1 Monoclonal Antibody (2B7)(Product # MA5-15815) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing mitochondrial localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of NTERA-2 cells using PRKAA1 monoclonal antibody (Product # MA5-15815) (Green). Blue: DRAQ5 fluorescent DNA dye. Red: actin filaments have been labeled with phalloidin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of NTERA-2 cells using PRKAA1 monoclonal antibody (Product # MA5-15815) (Green). Blue: DRAQ5 fluorescent DNA dye. Red: actin filaments have been labeled with phalloidin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of paraffin-embedded ovarian cancer (left) and brain tissues (right) using PRKAA1 monoclonal antibody (Product # MA5-15815) followed with DAB staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometric analysis of PC-2 cells using PRKAA1 monoclonal antibody (Product # MA5-15815) (green) and negative control (purple).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Basal alpha1AMPK regulates VE-Cad, ZO-1, and Cx43 expression. ( a ) Schematic representation of the experimental design, in which HMECs were transfected with control non-targeting siRNA or alpha1AMPK siRNA (50 nM) for 48 h. Then, they were treated with 991 (1 muM) or DMSO for one hour and subsequently exposed or not exposed to lipopolysaccharide (LPS, 50 muM) for six hours; ( b , c ) Human dermal microvascular endothelial cells (HMECs) were treated according to the protocol detailed in ( a ). Cell lysates were submitted to Western blot analysis and probed with alpha1AMPK, phospho-AMPK (Thr172), phospho-ACC (Ser79), VE-Cad, ZO-1, and Cx43 antibodies (Abs). Anti-eukaryotic elongation factor 2 (eEF2) was used as a loading control. Representative western blots; ( b ) and quantifications ( c ) are shown. Data are expressed as mean +- standard deviation (SD) (three biological replicates for each condition). # p < 0.05 is relative to corresponding untreated HMECs and * p < 0.05 is relative to cells transfected with the scrambled siRNA. The data underwent two-way analysis of variance (ANOVA).

MA5-15815

Antibody data