Question (59823) | Caspase-3 Assay Kit (Colorimetric) (ab39401)

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Question

Hello, Abcam Tech Support,   I would like to know bout the following issues for my purchase of this kit, Caspase 3 assay kit, ab39401.   1 Does this kit recognize rat caspase-3? 2 Does this kit recognize both activated and non-activated caspase-3 or either? 3 Can I use this kit for tissue caspase 3 assay? If so, a 50-200 mcg protein use remains same as a recommended protocol? 4 For tissue assay, any additional suggestions on the protocol?   Thank you in advance for your reply,  

Answer

Thank you for contacting us. I have answered each of your questions below. Please let me know if you have any additional questions about this or any Abcam product.

1. Does this kit recognize rat caspase-3?

This will recognize all mammalian forms (90% homology of Caspase 3 w/rat)



2. Does this kit recognize both activated and non-activated caspase-3 or either?

The assay is based on spectrophotometric detection of thechromophore p-nitroaniline (p-NA) after cleavage from the labeledsubstrate DEVD-p-NA. Therefore it should only recognize activated Caspase 3.



3. Can I use this kit for tissue Caspase 3 assay? If so, a 50-200 mcg protein use remains same as a recommended protocol?

Yes, you can use tissue lysates for this kit. Use exactly the same protocol. Use 200-400 µl of the lysis buffer for ach 10 mg of tissue. Samples should be completely homogenized, preferably using a Dounce homogenizer. Some substances can interfere with the assays and should be avoided in sample preparation. Please see individual kit datasheets for more specific information on which substances may affect the specific kits, but in general, substances such as EDTA (> 0.5mM), ascorbic acid (> 0.2%), sodium azide (> 0.2%), NP-40 and Tween-20 (>1%) are known to interfere with these assays and should be avoided in sample preparation. Moreover, we recommend deproteinating the sample prior analysis using a 10kD spin column (ab93349).



4. for tissue assay, any additional suggestions on the protocol?

Use exactly the same protocol. Use 200-400 µl of the lysis buffer for ach 10 mg of tissue.






I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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