Immunoblotting analyses of PrP relied on the monoclonal 3F4 antibody (catalog number MAB1562, Millipore Sigma) which recognizes the 109–112 amino acid epitope of human PrP. Antibodies against the NKA subunits consisted of anti-ATP1A1 (catalog number ab7671, Abcam Inc., Toronto, ON, Canada), anti-ATP1A2 (catalog number ab9094-I, Abcam Inc.), anti-ATP1A3 (catalog number MA3-915, Thermo Fisher Scientific) and anti-ATP1B1 (catalog number GTX113390, GeneTex, Irvine, CA, USA). Neuronal and astrocytic markers were detected with the anti-NeuN antibody (catalog number EPR12763, Abcam Inc.), the anti-Tuj-1 antibody (catalog number ab78078, Abcam Inc.) and the anti-GFAP monoclonal (131-17719) antibody (catalog number A-21282, Thermo Fisher Scientific).
Anti atp1a2
Manufactured by Abcam
Sourced in Canada, United Kingdom, China
Anti-ATP1A2 is a primary antibody that recognizes the ATP1A2 protein, which is the catalytic alpha subunit of the sodium-potassium ATPase (Na+/K+ ATPase) enzyme. The sodium-potassium ATPase is responsible for maintaining the electrochemical gradients of sodium and potassium ions across the cell membrane.
Automatically generated - may contain errors
Lab products found in correlation
Anti gfap, by Thermo Fisher Scientific (1 mentions)
Anti atp1a1, by Abcam (1 mentions)
Anti tuj1 antibody, by Abcam (1 mentions)
Anti neun antibody, by Abcam (1 mentions)
Enhanced chemiluminescence reagent, by Beyotime (1 mentions)
Matrix metalloproteinase mmp 2, by Proteintech (1 mentions)
E cadherin, by Proteintech (1 mentions)
Anti n cadherin, by Cell Signaling Technology (1 mentions)
Anti β catenin, by Cell Signaling Technology (1 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Anti snail, by Proteintech (1 mentions)
α sma, by Cell Signaling Technology (1 mentions)
Anti tgf β, by Cell Signaling Technology (1 mentions)
Anti vimentin, by Cell Signaling Technology (1 mentions)
Anti smad2, by Cell Signaling Technology (1 mentions)
Anti smad3, by Cell Signaling Technology (1 mentions)
Anti p smad2, by Cell Signaling Technology (1 mentions)
Anti p smad3, by Cell Signaling Technology (1 mentions)
β actin, by Proteintech (1 mentions)
Caspase 3, by Proteintech (1 mentions)
3 protocols using anti atp1a2
1
Immunoblotting Analysis of Prion Proteins
2
Comprehensive Western Blot Protein Analysis
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Western blot was performed as previously described (20 (link),21 (link)). Cell proteins were extracted with RIPA lysis buffer (KeyGene Biotech, Nanjing, China), and the protein concentration was determined by BCA (KeyGene Biotech). After SDS-PAGE electrophoresis and membrane transfer, the membranes were blocked with 5% non-fat milk for 1 h and then incubated with primary antibody overnight at 4 °C. After incubation with horseradish peroxidase-conjugated secondary antibodies at room temperature for 1 h, the membranes were visualized using enhanced chemiluminescence reagents (Beyotime). The primary antibodies used in the study included Bax, caspase-3, Bcl-2, E-cadherin, matrix metalloproteinase (MMP)-2, MMP-9, and β-actin (purchased from ProteinTech, IL, USA), anti-Snail, anti-vimentin, anti-N-cadherin, anti-β-catenin, anti-TGF-β, α-SMA, anti-p-Smad2, anti-Smad2, anti-p-Smad3, anti-Smad3, and anti-cleaved caspase-3 were purchased from Cell Signaling Technology (MA, USA). Anti-ATP1A2 was supplied by Abcam (Cambridge, UK).
3
Plasma Membrane Protein Isolation and Analysis
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Intestinal tissue and IEC-6 cells membrane protein was isolated by MinuteTM Plasma Membrane Protein Isolation Kit (Invent Biotechnologies, Inc. Plymouth, MN, USA) [44 (link)]. Total proteins of intestinal tissue and IEC-6 cells were extracted in lysis buffer (25 mM Tris-HCl pH7.4, 150 mM NaCl, 1% NP-40, 1 mM EDTA, 5% glycerol, 0.1% SDS) containing protease and phosphatase inhibitors. Then, 20 μg of protein were subjected to SDS-PAGE and transferred to PVDF membranes for immune-blot analysis. The following antibodies were used: GLUT2 (Santa Cruz, SC-9117, 1:500, Dallas, TX, USA), GLUT5 (Santa Cruz, SC-30109, 1:500, Dallas, TX, USA), SGLT1 (Abcam, ab14686, 1:500, Waltham, MA, USA), PLC-β2 (Santa Cruz, SC-206, 1:500, Dallas, TX, USA), IGF-1R-β (Cell Signaling Technologies, 9750S, 1:1000, Shanghai, China), phospho-IGF-1R-β (Tyr 1135) (Cell Signaling Technologies, 3918, 1:1000, Shanghai, China), β-Actin (Cell Signaling Technologies, 3700S, 1:1000, Shanghai, China), Anti-ATP1A2 (Abcam, ab2871, 1:1000, Waltham, MA, USA). The intensity of the specific immune-reactive bands was quantified using Quantity One software (Bio-Rad Laboratories, Inc. Hercules, CA, USA).
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