For in vitro acetylation reaction, 1 µg recombinant KAT7 were incubated with 5 µg recombinant vimentin, 10 µg polypeptide of vimentin‐WT (NTRTNEKVELQELC) or 1 µg polypeptide of vimentin‐K104Ac (NTRTNE‐Acetyl‐K‐VELQELC) in the HAT buffer containing 2 mm acetyl‐coenzyme A (10101893001, Roche, Switzerland), 50 mm pH 8.0 Tris–HCl (ST780, Beyotime, Shanghai, China), 50 mm NaCl, 4 mm MgCl2, 0.1 mM EDTA (C0196, Beyotime, Shanghai, China), 1 mm DTT (ST040, Beyotime, Shanghai, China), and 10% glycerol (ST1348, Beyotime, Shanghai, China) at 37 °C for 1 h. Loading buffer (2×) was added in the above reaction system, and acetylation of vimentin was detected using western blot with an anti‐vimentin‐K104Ac antibody.
St040
Manufactured by Beyotime
Sourced in China, United States
The ST040 is a laboratory equipment product. It is designed to serve a core function, but a detailed description while maintaining an unbiased and factual approach is not available.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using st040
1
Vimentin Acetylation by KAT7 In Vitro
2
Colorimetric Assay for Calcineurin
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Lyophilized CNAΔ316, CNB standard, CNB test sample (Beijing Normal University), p-nitrophenylphosphate (pNPP) (34047, Thermo Fisher Science, USA), bovine serum albumin (BSA) (ST023 Beyotime, Shanghai), DLdithiothreitol (DTT) (ST040, Beyotime, Shanghai), and anti-CNB polyclonal antibody (ab154650, Abcam, USA) were used.
Testing medium comprised 50 mmol L 1 Tris-HCl (pH 7.4), 0.2 mg mL 1 BSA, 20 mmol L 1 pNPP, 1 mmol L 1 DTT, 1 mmol L 1 MnCl 2 , and 1 mmol L 1 CaCl 2 . Microtiter plate reader (Spectrum plus 5, Molecular Device, USA) was used in the assays.
Testing medium comprised 50 mmol L 1 Tris-HCl (pH 7.4), 0.2 mg mL 1 BSA, 20 mmol L 1 pNPP, 1 mmol L 1 DTT, 1 mmol L 1 MnCl 2 , and 1 mmol L 1 CaCl 2 . Microtiter plate reader (Spectrum plus 5, Molecular Device, USA) was used in the assays.
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