All cellular proteins were extracted using lysis buffer (Beyotime, China) with phosphatase inhibitor (Roche, Basel, Switzerland). The proteins were loaded in SDS-PAGE to be separated and transferred to PVDF membranes (Millipore, Burlington, MA, USA). The PVDF membranes were blocked with 10% skimmed milk in TBST and incubated with the primary antibodies specific for a rabbit anti SPC21 antibody at 1:500 (Novusbio Biologicals, Hubei, China), a rabbit anti PTEN antibody at 1:800 (Abcam, Waltham, MA, USA), a rabbit anti p-Akt antibody at 1:1000 (CST, USA), a rabbit anti Akt antibody at 1:1000 (CST, USA), and a rabbit anti β-actin antibody at 1:2000 (Abcam). After being washed with TBST three times for 15 min, the membranes were incubated with secondary antibodies anti-rabbit or anti-mouse IgG biotinylated at 1:10000 (Solarbio, Beijing, China) for 2 h at room temperature. Protein bands were visualized by the ECL Chemiluminescent Kit (Millipore) and a ChemiDoc Touch imaging system (Bio-Rad, Hercules, CA, USA).
Rabbit anti pten antibody
Manufactured by Abcam
Sourced in China, United Kingdom, United States
Rabbit anti-PTEN antibody is a primary antibody that recognizes the PTEN protein, a tumor suppressor that regulates cell growth and division. This antibody is useful for the detection and analysis of PTEN expression in various applications, such as Western blotting, immunohistochemistry, and immunocytochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (1 mentions)
Chemidoc touch imaging system, by Bio-Rad (1 mentions)
Phosphatase inhibitor, by Roche (1 mentions)
Rabbit anti β actin antibody, by Abcam (1 mentions)
Lysis buffer, by Beyotime (1 mentions)
Ecl chemiluminescent kit, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Pvdf membrane, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Abcam (1 mentions)
Rabbit anti pdcd4 antibody, by Abcam (1 mentions)
Protein estimation kit, by Merck Group (1 mentions)
β actin, by Abcam (1 mentions)
3 protocols using rabbit anti pten antibody
1
Western Blot Analysis of Cellular Proteins
2
PTEN Expression Quantification in FLS
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PTEN expression in FLS treated with TNF-α was detected by immunocytochemistry using a rabbit anti-PTEN antibody (Abcam, Cambridge, United Kingdom). The cells were stained with 4′,6-diamidino-2-phenylindole (Beyotime, Shanghai, China) in the dark and then photographed under an epifluorescence microscope (BX-51; Olympus, Tokyo, Japan).
3
Western Blot Analysis of Spinal Cord Injury
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The animals were subjected to euthanasia by overdose of sodium pentobarbital (240 mg/ml) divided into day 1 and 3 after injury. The spinal cords were harvested and a segment measuring 1 cm with the injured site was used for studying the expression of proteins by western blot. The harvested spinal tissues were homogenized and subjected to lysis, and total protein was determined using protein estimation kit (Sigma Aldrich USA). About 20 µg of protein was submitted to electrophoresis on sodium dodecyl sulfate/polyacrylamide gel (12%) and then transferred to PVDF membranes (Thermo Fisher USA). The PVDF membranes were blocked using 5% nonfat milk for 1 h. The membranes were incubated for 12 h at 4ºC along with rabbit anti-FasL antibody (1 : 1000, Abcam USA), rabbit anti-PTEN antibody (1 : 1000, Abcam USA), rabbit anti-PDCD4 antibody (1 : 1000 Abcam USA) and β-actin (1 : 1000, Abcam, USA). The membranes were then incubated with horseradish peroxidase conjugated IIry antibody (1 : 10000) for 60 min at 37ºC. The proteins were viewed using enhanced chemiluminescence; β-actin was used as loading control.
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