Caspase 1 Assay Kit (Fluorometric) (ab39412)

What is the range of sensitivity of this assay?

ANSWER:

Unfortunately we do not have a sensitivity range determined for this kit.

Fold-increase in Caspase-1 activity can be determined by comparing the results of induced samples with the level of the untreated control.

Please let me know if you have any more questions. 

I'm using mouse left ventricle with this kit. I was wondering what type of fold increase has been seen before when testing this kit?

ANSWER:

Thank you for your inquiry.

This result would depend on a number of factors including the cell type in which the Capase-1 activation is being studied, the levels of Caspase-1 activation after induction of apoptosis, time-course for Caspase-1 activation after apoptosis induction. We do have the following citations for this kit which will be of help to you to give you an idea:

Caspase-1 Fluorometric Assay Kit

Cheng, S. C. et. al. The dectin-1/inflammasome pathway is responsible for the induction of protective T-helper 17 responses that discriminate between yeasts and hyphae of Candida albicans. J. Leukoc. Biol., Aug 2011; 90: 357 - 366.

Cihakova D et al (2008) Am. J. Pathol. 172: 1195-1208.

Coletti, D., et al. (2002) EMBO J. 21:631-642.

Duncan JA et al (2009) J. Immunol.; 182: 6460-6469Koenen, T. b. et. al. Hyperglycemia Activates Caspase-1 and TXNIP-Mediated IL-1β Transcription in Human Adipose Tissue. Diabetes, Feb 2011; 60: 517 - 524. 

I hope this information helps. Please contact us with any other questions.

How should tissue be prepared to use with the kit? Any specific protocol?

After lysing tissue in the cell lysis buffer, can I use this tissue lysate in a Western blot?

ANSWER:

Thank you for your call today. I have heard back from the lab about your questions:

1) For tissues, homogenize 10 mg of tissue in 200 ul of lysis buffer. Spin out debris and add 50 ul of supernatant into a well of the 96-well plate then proceed from Step 4 on the protocol.

2) You can use the lysate for Western blot.

I hope this information is helpful, but please let me know if you have any further questions.

Products ab39470 and ab39412. What sensitivity (pg, ng, ug?) do these products range from, for the detection of Caspase1?

ANSWER:

Thank you for your enquiry.

The colorimetric assay has sensitivity of about 5 ng and fluorometric is about 10-100 fold more sensitive. Fluorometric assay is recommended for tissue samples.

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