乙酰化蛋白快速检测和定量
Rapid detection and quantitation of total cellular acetylated proteins using our research products
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Illustration of the principle of the methods:
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Procedures:
1) 400 ug of crude proteins from cells of each treatment group was enriched with 10 mL of the anti-AcK Agarose (ICP0388).
2) The bound acetylated species were eluted with 100 mL of 0.1 M HCl twice.
3) The eluates were pooled and pH was adjusted with 10 mL of the saturated Na2CO3.
4) 100 uL/well was added to 8-well strip plates in duplicate and was incubated overnight at room temperature.
5) The strip plate was then blocked with 0.1% BSA in PBSt and washed three times with PBSt.
5) Anti-AcK HRP (ICP0381) (0.25 ug/mL in 0.1% BSA in PBSt) was added at 37oC for 30 min.
6) The strip plate was washed three times with PBSt and100 uL of TMB was added and incubated for 30 minutes to allow for color development.
7) The colour reaction was impeded by the addition 10 uL of 2M H2SO4. Read absorbance at 450 nm.
8) The reactive acetylated protein level was caculated by OD (treatment)/OD (no-treatment).
One Study: Analyisis of total acetylated protein level in cells treated with various concentrations of TSA for 12 hours.
Increased protein acetylation up to 10 folds in cells treated with different concentration of TSA using the IP-ELISA combined method (IP with anti-AcK Agarose (ICP0388), detected with anti-AcK HRP (ICP0381).
Western blot analysis of acetylated proteins from samples of MMRU cell treated with HDAC inhibitor TSA for 5 and 15 hours respectively.
The sample was affinity-enriched with anti-acetylated lysine agarose (ICP0388).
Lane 1&2: Western-blot (WB) with anti-acetylated lysine antibody HRP conjugated (ICP0381).
Lane 3: Molecular maker
Lane 4&5: Western-blot (WB) with anti-acetylated lysine antibody HRP conjugated (ICP0381) + 10mg/mL acetylated BSA (ICP6090).
| Related Research Tools |
| Acetyl Lysine Antibody - ICP0380 It recognizes proteins with acetylated lysine. For ELISA, WB, IP, immunofluorescence, and immunochemistry. 100ug - $350 Acetyl Lysine Antibody, Mouse Monoclonal - ICP0390 It recognizes proteins acetylated on lysine residues. For ELISA, WB, IF, and IP 100ug - $350 |
| Acetyl Lysine Antibody, HRP - ICP0381 It recognizes proteins acetylated on lysine residues. For direct immunofluorescence experiments. 100ug - $395 Acetyl Lysine Antibody - FITC - ICP0385 It recognizes proteins acetylated on lysine residues. For direct immunofluorescence experiments. 100ug - $375 |
| Acetyl Lysine Antibody, Biotin - ICP0383 It recognizes proteins acetylated on lysine residues. 100ug - $375 Acetyl Lysine Antibody, Agarose - ICP0388 It selectively captures peptides and proteins with acetylated lysine residues (N-epsilon). For rapid isolation and purification of peptides or proteins with acetylated lysine residues from cell lysate or protease digested mixtures 2mg - $550 5mg - $950 |
| Acetylated BSA - ICP6090 It is well recognized and binds strongly to affinity purified anti-acetylated lysine antibodies. Positive control and competitive inhibitor for anti-acetylated lysine antibodies. 5mg - $295 With delivery times of 1-3 business days, what are you waiting for? Contact us orders@immunechem.comfor quotes on bulk orders! |
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