For kit PF0100, it's recommended to transfer the samples from the centrifuge to a secondary container before pipetting into the cuvette or well plate to ensure homogeneity and prevent false positive readouts during fluorescent detection.
PF0100
Protease Fluorescent Detection Kit
high sensitivity assay
Synonym(s):
Protease Assay Kit
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About This Item
UNSPSC Code:
12352202
NACRES:
NA.26
Pricing and availability is not currently available.
Recommended Products
Application
Protease Fluorescent Detection Kit has been used:
- in the detection of proteolytic activity using casein based protease assay[1]
- for testing the extracellular proteolytic activity of fungal strains[2]
- for quantifying mitochondrial protease activity[3]
- for measuring the protease production from soil solution[4]
Suitability
Detection limit: approx. 50 ng trypsin
Quantity
Supplied with sufficient reagents for up to 200 one mL assays.
Supplied with sufficient reagents for up to 200 one mL assays.
The trichloroacetic acid (TCA) component is provided as 6.1 N TCA solution. 0.6 N TCA Solutions may be prepared by a 10-fold dilution of the desired volume of T0699 with ultrapure water. It is not necessary to dilute the entire volume of T0699 for immediate use of the kit.
The trichloroacetic acid (TCA) component is provided as 6.1 N TCA solution. 0.6 N TCA Solutions may be prepared by a 10-fold dilution of the desired volume of T0699 with ultrapure water. It is not necessary to dilute the entire volume of T0699 for immediate use of the kit.
Analysis Note
Tested for detection of all four proteolytic classes (serine, apartic, cysteine and metalloproteinases).
Kit Components Only
Product No.
Description
- Incubation buffer 25 mL
- FITC-casein solution 5 mL
- Assay buffer 200 mL
- Protease standard (trypsin) 20 μg
- FITC control 50 mg
- Trichloroacetic acid (TCA) solution 30 mL
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Aquatic Acute 1 - Aquatic Chronic 1 - Eye Dam. 1 - Resp. Sens. 1 - Skin Corr. 1A - Skin Sens. 1 - STOT SE 3
Target Organs
Respiratory system
Storage Class Code
8A - Combustible corrosive hazardous materials
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
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Articles
Get better detection and quantification of proteases with this high-sensitivity red protease detection assay.
Get better detection and quantification of proteases with this high-sensitivity red protease detection assay.
Get better detection and quantification of proteases with this high-sensitivity red protease detection assay.
Get better detection and quantification of proteases with this high-sensitivity red protease detection assay.
Protocols
Protease Fluorescent Detection Kit detects protease activity using FITC-labeled casein substrate.
Protease Fluorescent Detection Kit detects protease activity using FITC-labeled casein substrate.
Protease Fluorescent Detection Kit detects protease activity using FITC-labeled casein substrate.
Protease Fluorescent Detection Kit detects protease activity using FITC-labeled casein substrate.
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Seeking clarity on step 7 on the procedure. Should the supernatant be collected? How should it be done without capturing any precipitate/pellet? This seems to be affecting themeasurements.
1 answer-
Helpful?
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If RIPA lysis buffer is used for cell lysis in cultured cells without isolating the mitochondria, will it have an impact on the protease detection when utilizing the PF0100-1KT Protease Fluorescent Detection Kit?
1 answer-
The QC tests for protease in the R0278 RIPA lysis buffer show no increase in background from the data, indicating no additional breakdown of the FITC-casein substrate when mixed with RIPA buffer. It is recommended that the use of R0278 or an equivalent, as this buffer is protease-free.
Helpful?
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I want to use the protease fluorescent detection kit in cultured cells without isolating the mitochondria. If I use RIPA lysis buffer for cell lysis, will it affect the protease detection?
1 answer-
RIPA lysis buffer is compatible with this kit, provided that it is protease free. Please see the link below to review product R0278, a protease free RIPA buffer option:
https://www.sigmaaldrich.com/CA/en/product/sigma/r0278Helpful?
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