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Caspase 6 Activity Assay Kit

Caspase 6 Activity Assay Kit

Catalog Number: abs50027 Brand: Absin

Price:

$123.00 USD

$123.00 USD

Size: 20T 100T

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Usage	Determination of pNA standard curve: 1. The 10 mM pNA standard was diluted with reaction buffer to 200, 100, 50, 25, 12.5, 6.25, 3.125 μM. Set the zero tube without pNA. 2. Take 100μl of each concentration and add it to a 96-well plate or 100μl cuvette, and measure the absorbance OD value at 405nm. 3. The OD405 value of each concentration standard tube is the x-axis, and the corresponding pNA concentration is the y-axis. Use Excel to make a standard curve of pNA concentration versus OD405 value. Detection of Caspase 6 enzyme activity: 1. Take out an appropriate amount of Ac-VEID-pNA (2mM) and place it on an ice bath for later use. 2. According to the 100ul reaction system: blank control group: 40μl of detection buffer, 0μl of sample to be tested, 50μl of lysate, 10μl of Ac-VEID-pNA (2mM); Sample group: 40μl of detection buffer, 50μl of sample to be tested, 0μl of lysate, 10μl3 of Ac-VEID-pNA (2mM). Add Ac-VEID-pNA (2mM) and mix well. Be careful to avoid bubbles during mixing. Incubate at 37 º C for 60-120 minutes. A405 can be measured when the color change is obvious. If the color change is not obvious, the incubation time can be appropriately extended, or even overnight. 4. The A405 of the sample minus the A405 of the blank control is the absorbance produced by pNA catalyzed by caspase 6 in the sample. By comparing the standard curves obtained in the above steps, it is possible to calculate the amount of pNA catalytically produced in the sample.
Theory	Based on Caspase 6, it can catalyze the substrate Val Glu Ile Asp p nitroanilide (Ac VEID pNA) to produce yellow pNA (p-nitroanilide), which can be used to detect the activity of Caspase 6 by measuring absorbance. pNA has a maximum absorbance at 405 nm. Caspase activity was obtained by measuring pNA using visible light colorimetry. Suitable for detecting Caspase 6 activity in mammalian tissues and cells.
Experiment Reagent Instrument	Microplate reader, spectrophotometer
Synonym	Mch-2,caspase-6,caspase 6
Description	caspase 6 Activity Detection Kit is a kit that detects caspase 6 enzyme activity or purified caspase 6 enzyme activity in cell or tissue lysates by spectrophotometry. Caspases are a family of proteases that play an important role in the process of apoptosis. The precursor of caspase 6 can form activated caspase 6 dimer after cleavage by granzyme B, and activated caspase 6 has been found to induce apoptosis. Caspase 6 can cleave PARP and keratin-18, as well as Lamin A, a key constituent protein on the nuclear envelope of the cell nucleus. Only caspase 6 of the Caspase family can cut Lamin A. The recognition site of Caspase 6 for Lamin A is VEID. This kit utilizes the specificity of caspase 6 for VEID recognition, and designs the corresponding caspase 6-specific chromogenic substrate Ac-VEID-pNA.