For analysis of alpha toxin binding and heptamerization, keratinocytes were treated with alpha toxin for two hours, washed with PBS, and harvested in hypotonic lysis buffer with 1% Triton X-100, containing protease inhibitor (Complete, Roche). Cellular debris was pelleted by centrifugation, clarified lysates were resuspended in Laemmli buffer and proteins resolved on a 5–15% gradient gel (Biorad). (The alpha-toxin heptamer is stable during electrophoresis.) Transferred proteins were blotted with anti-alpha toxin antibody (Sigma), and detected by enhanced chemiluminescence (Amersham). Scanned images were quantitated with Image J software. For immunoblotting Actin (Sigma), ADAM10 (Millipore), phospho-STAT1 (Cell Signaling) and LC3 (Abcam), cells were harvested, lysed and probed as described above.
Anti alpha toxin antibody
Manufactured by Merck Group
The Anti-alpha toxin antibody is a laboratory reagent used for the detection and quantification of alpha toxin, a virulence factor produced by certain bacteria. This antibody can be utilized in various immunoassay techniques to identify and measure the presence of alpha toxin in samples.
Automatically generated - may contain errors
4 protocols using anti alpha toxin antibody
1
Analysis of Alpha Toxin Binding and Heptamerization
2
Analysis of Alpha Toxin Binding and Heptamerization
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For analysis of alpha toxin binding and heptamerization, keratinocytes were treated with alpha toxin for two hours, washed with PBS, and harvested in hypotonic lysis buffer with 1% Triton X-100, containing protease inhibitor (Complete, Roche). Cellular debris was pelleted by centrifugation, clarified lysates were resuspended in Laemmli buffer and proteins resolved on a 5–15% gradient gel (Biorad). (The alpha-toxin heptamer is stable during electrophoresis.) Transferred proteins were blotted with anti-alpha toxin antibody (Sigma), and detected by enhanced chemiluminescence (Amersham). Scanned images were quantitated with Image J software. For immunoblotting Actin (Sigma), ADAM10 (Millipore), phospho-STAT1 (Cell Signaling) and LC3 (Abcam), cells were harvested, lysed and probed as described above.
3
Alpha Toxin Binding and Heptamerization
4
Alpha Toxin Binding and Heptamerization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For analysis of alpha toxin binding and heptamerization, keratinocytes were treated with alpha toxin for one hour, washed with PBS, and harvested (without trypsin) in hypotonic lysis buffer with 1% Triton X-100, containing protease inhibitor (Complete, Roche). Cellular debris was pelleted by centrifugation and clarified lysates were resuspended in Laemmli buffer and proteins resolved on a 5-15% gradient gel (Biorad). (The alpha-toxin heptamer is stable during electrophoreses.) Transferred proteins were blotted with anti-alpha toxin antibody (Sigma), and detected by enhanced chemiluminescence (Amersham). Scanned images were quantitated with Image J software. For immunoblotting ADAM10 (antibody from Millipore), cells were harvested, lysed and probed as described above, but without alpha toxin treatment.
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