mTOR Polyclonal Antibody Detects Target Bands in HeLa and Jurkat Cell Lines
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The customer review summarizes results from testing St John's Laboratory's mTOR Polyclonal Antibody (STJ94280) via western blot. The antibody detected target bands at the expected size of 288kD in HeLa and Jurkat cell lysates. The protocol details the cell lysis, gel electrophoresis, transfer, blocking, primary and secondary antibody probing, washing, and visualization steps. The customer stated that the testing results provided by the testing laboratory showed the target bands were detected by the antibody in the two cell lines. St John's Laboratory invites customers to test their products for free by applying for trial samples online.
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mTOR Polyclonal Antibody Detects Target Bands in HeLa and Jurkat Cell Lines
- 1. HOW DID OUR ANTIBODY PERFORM? CUSTOMER REVIEW: mTOR Polyclonal Antibody (STJ94280)
- 2. WHAT WAS USED? Primary antibody: STJ94280. mTOR Polyclonal Antibody. 100µl Provider: St John's Laboratory Dilution ratio: 1:1000 Application: Western Blot. Image: In HeLa and Jurkat cell line, there were target bands detected by STJ94280 mTOR Polyclonal Antibody.
- 3. WHAT WAS THE PROTOCOL? Treatment of materials: When cells were cultured to 70%-80% plating density, lysis buffer with protease inhibitors (Sigma) and equal volume of 2X loading buffer were added for 10 min water bath at 100˚C. Gel electrophoresis: 10% polyacrylamide gel, constant voltage 160 V, 60 min. Transfer: 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min. Blocking: 1X TBST with 5% skimmed milk powder for 2 hr. Primary antibody probing: primary antibody was added to the solution of 1X TBST with 5% skimmed milk powder, 4˚C for overnight. Membrane wash: 1X TBST wash for 3 times. Secondary antibody probing: secondary antibody was added to the solution of 1X TBST with 5% skimmed milk powder with 1:10000 dilution ratio, for 1.5 hr. Membrane wash: 1X TBST wash for 3 times. Visualization: ECL visualization for 5 min (the visualization time was determined by actual result. It was ensured that the visualization time was identical for the same antigen).
- 4. WHAT WAS THE PROTOCOL? Gel electrophoresis information：10% polyacrylamide gel, constant voltage 160 V, 60 min. Transfer information：0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min. No. Antigen Loading amount Primary antibody Primary antibody dilution ratio Secondary antibody dilution ratio Target band KD Visualization time 1 HeLa lysate 10 µg STJ94280. mTOR Polyclonal Antibody 1:1000 1:10000 288KD 35S 2 Jurkat lysate 10 µg 1:10000 288KD 35S
- 5. WHAT DID THE CUSTOMER THINK? Testing results were provided by BPI laboratory. They stated that “In HeLa and Jurkat cell line, there were target bands detected by STJ94280 mTOR Polyclonal Antibody”.
- 6. “WE INVITE YOU TO TEST OUR PRODUCTS” Our aim is to not only connect our customers with the best product, but to also give them the opportunity to test the product’s performance risk free.
- 7. Contact Us: info@stjohnslabs.com 1. Choose 5 Trial Size Samples from our free sample list 2. Complete the free sample application: http://www.stjohnslabs.com/free-sample 3. Receive your antibody 4. Submit your review 5. Receive your 30% off your next order Get Your Free Samples in 5 Easy Steps
- 8. Follow Us for Weekly Independent Validation Reviews @StJohnsLabsSt John's Laboratory Ltd St John's Laboratory Ltd St John's Laboratory Ltd http://www.slideshare.net/stjohnslabs