Caspase 1 Assay Kit (Colorimetric) (ab39470)

I am sorry to trouble you. I would like to buy two Elisa-Kit to detect 1.mature mouse IL-1β and 2.mature mouse caspase-1 ，and I don't want it have a corssreactivity with pro-IL-1β and pro- caspase-1 , I found your company have these kit, could you tell me ,is this kit can only detect mature protein?
Thank you so much ,appreciate˜

ANSWER:

Thank you for contacting Abcam.

For ab39470, it measures the mature form of Caspase-1 and for ab100705, IL1 beta Mouse ELISA Kit - this kit will recognize the mature form (aa 118-269) of IL1beta. Since the pro IL1beta also contains this amino acid sequence, I would expect the kit to also detect the pro-form.

If there is anything else I can help you with, please let me know.

Phone call requesting information on whether mouse, human or both samples can be used with this kit.

ANSWER:

Thank you for contacting us and your interest in our products.


The Caspase 1 Colorimetric Assay Kit has been tested in house with a human cell line (Jurkat cells), but the kitshould work for almost all mammalian cells. We have several reports from other customers who have used this kit with mouse samples successfully


I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

in which step is the dilution buffer to be used? it is not mentioned in the protocol

ANSWER:

Thank you for contacting us. I have heard back from the lab with the following information: The dilution buffer can be used to dilute the caspase 1 protein (ab39901) when being used as a positive control. Thus, the protocol is correct as it is.

Could you please describe the problem you encountered as well as the type of samples you have been using? Were there any deviations from the protocol that you might have implemented? What is the order or PO number used to purchase the kit?

Please let me know this information so that I can help you further with resolvng the issues with this product. I look forward to hear back from you.

Answer to your queries,

1. No we have not checked the cells under microscope whether they are lysed or not, we can check that in our next set of experiments.

2. We are using 500,000 synoviocytes per well with or without treatments and let them grow till confluence, then we are collecting the cells and lysing them as per your protocol. For protein concentration, we are using 100ug of protein

in 50ul of lysis

buffer (protein concentration is estimated by BCA assay)

3. We are using freshly thawed DTT solution for our assays

4. The plate was read at 405nm wavelength.

Thank you for your reply. I will definitely get back once we get the results of our next set of experiments with this kit.

ANSWER:

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 953886.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee.

The reference that I mentioned for staurosporine induction of caspase-dependent apoptosis is:

Staurosporine induces apoptosis through both caspase-dependent and caspase-independent mechanisms. Oncogene. 2001 Jun 7;20(26):3354-62.

http://www.ncbi.nlm.nih.gov/pubmed/11423986

For a protocol specific to synoviocytes, the following may be of interest:

Angiotensin II protects fibroblast-like synoviocytes from apoptosis via the AT1-NF-κB pathway. Rheumatology (Oxford). 2007 Aug;46(8):1252-7. PMID:17526929

http://rheumatology.oxfordjournals.org/content/46/8/1252.long

I hope these are useful. Please contact us if you have any questions or concerns.

Answer to your queries,

1. No we have not checked the cells under microscope whether they are lysed or not, we can check that in our next set of experiments.

2. We are using 500,000 synoviocytes per well with or without treatments and let them grow till confluence, then we are collecting the cells and lysing them as per your protocol. For protein concentration, we are using 100ug of protein

in 50ul of lysis

buffer (protein concentration is estimated by BCA assay)

3. We are using freshly thawed DTT solution for our assays

4. The plate was read at 405nm wavelength.

Thank you for your reply. I will definitely get back once we get the results of our next set of experiments with this kit.

ANSWER:

Thank you for providing this additional information. Please let me know how your next set of experiments goes and if you are not able to improve your results, I would be happy to offer you a free of charge replacement, credit, or refund.