A431 + Calyculin A (30min) lysate
€155.00
In stock
SKU
ECM-AL9101
Background:
Calyculin A is a serine/threonine phosphatase inhibitor that inhibits the activity of protein phosphatases PP1 and PP2A. Human carcinoma A431 cells treated with calyculin A for 30 minutes can undergo significant threonine phosphorylation, as shown by western blotting using anti-Phospho-Akt (Thr-34), cat.# AP1001, as compared to untreated, control cell lysates. Confluent cultures of A431 cells were serum starved overnight. Cells were then either left untreated (Cat.# AL9001) or treated with Calyculin A at a final concentration of 100 nM for 30 minutes at 37°C (Cat.# AL9101). Cells were lysed in 1% SDS, 1.0 mM sodium ortho-vanadate, 1 mM sodium fluoride, 10 mM Tris (pH 7.4) buffer. Protein concentration was determined using the BCA method (Pierce) before diluting to final concentration and buffer.
Application dilution:
WB: 20 μl/lane
End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1 hour at room temperature.
Buffer/Storage:
Cell Lysates are supplied at a concentration of 1 mg/ml in electrophoresis sample buffer (62.5 mM Tris pH 6.8, 2% SDS, 5% glycerol, 0.003% bromophenol blue, 0.9% β-mercaptoethanol). Store at –20°C. Do not boil or dilute. Stable for 1 year.
Calyculin A is a serine/threonine phosphatase inhibitor that inhibits the activity of protein phosphatases PP1 and PP2A. Human carcinoma A431 cells treated with calyculin A for 30 minutes can undergo significant threonine phosphorylation, as shown by western blotting using anti-Phospho-Akt (Thr-34), cat.# AP1001, as compared to untreated, control cell lysates. Confluent cultures of A431 cells were serum starved overnight. Cells were then either left untreated (Cat.# AL9001) or treated with Calyculin A at a final concentration of 100 nM for 30 minutes at 37°C (Cat.# AL9101). Cells were lysed in 1% SDS, 1.0 mM sodium ortho-vanadate, 1 mM sodium fluoride, 10 mM Tris (pH 7.4) buffer. Protein concentration was determined using the BCA method (Pierce) before diluting to final concentration and buffer.
Application dilution:
WB: 20 μl/lane
End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1 hour at room temperature.
Buffer/Storage:
Cell Lysates are supplied at a concentration of 1 mg/ml in electrophoresis sample buffer (62.5 mM Tris pH 6.8, 2% SDS, 5% glycerol, 0.003% bromophenol blue, 0.9% β-mercaptoethanol). Store at –20°C. Do not boil or dilute. Stable for 1 year.
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