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Glutathione Assay Kit (Fluorescence)

Catalog Number:
CA-G055
Price: $249.00
Detailed Description

CA-G055

Name

EliteTM Glutathione (GSH) Assay Kit (Fluorescence)

Description

The EliteTM Glutathione Assay Kit provides an ultrasensitive fluorimetric assay to quantify GSH in sample. The proprietary non-fluorescent glutathione sensor used in the kit becomes strongly green fluorescent upon reacting with a GSH compound, which has the spectral properties almost identical to those of fluorescein and can be easily read by a fluorescence microplate reader at Ex/Em = 490/520 nm. The kit can detect as little as 1 picomole of GSH in a 100 μL assay volume (10 nM). In addition, both absorption and emission spectra of the glutathione adduct are pH-independent, making this assay kit highly robust.

Application

The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step.

Size

200 assays 

Detection

Fluorescence microplate reader

 

Click here for DATA SHEET

Click here for MSDS




Frequently Asked Questions:


1. Q: What would you suggest to prepare the cell samples for this assay?

Answer: All the detergents in various lysis buffers will cause background noise for the assay. The traditional way to do it is the "freeze and thaw" method (i.e. freeze for 20-30 minutes at -80 C then thaw at room temperature, 2-3 times), but it is too tedious. We found that 0.2% SDS gives the best results among all the detergents tested.


2. Q: Does this Kit work for plant samples? Could you recommend a special protocol to measure gluthatione in plants? Answer: As long as the sampl chop 2 g of plant parts (leaf, root, grain, etc.) and ground it in a homogenizer for 2 min with 5-10 ml of 0.1 N HCl; centrifuge to pbtain the GSH containing supernatant, then adjust pH to 6.5 for the GSH assay.


3. Q: My sample is frozen brain tissue, how should I prepare it for the saasy?

Answer: It is better use PBS, and simply homogenize it, then use the supernatant.


4. Q: I work with human primary cells. How many cells should I use in the assay?

Answer: Suggest to start with 10,000 cells/well first.


5. Q: For the fluorescent detection, an emission filter with 520 nm is recommended. But we have only 535 nm filter. Would it work for this measurement?

Answer: Yes.


6. Q: We don't have the filters for 490/520 (nm). What we have are for 395/535 (nm). Would it work?

Answer: No. the emission is OK, but the excitation is not. The lowest excitation wavelength is at 460 nm.


7. Q:  Could you take a look at my protocol for preparing the cell culture lysate for the assay and give your suggestion?

Answer:

    - remove supernatant, then wash with HEPES-BSS;[Fine]

    - trypsinize for 2-6 min, stop trypsination;[We suggest to avoid trypsin and try using a rubber scrapper or EDTA to harvest cells, then centrifuge the cells and use the supernatant]

    - centrifuge pellet down at 180g, 5min, 4°C, then wash with 5ml cold PBS; count cells; [Fine]

    - centrifuge pellet down at 180g, 5min, 4°C; [May need to use more cells since you indicated the signal in the undiluted sample is very low.]

    - wash with 5ml COLD PBS; resuspend in ice-cold 5% MPA;[Fine. 5% MPA could be added to the supernatant for deproteination.]

    - mix thoroughly, homogenize by sonication (2x 5 cycles), then centrifuge at 12.00rpm for 5min at 4°C;[Fine]

    - collect supernatant for GSH assay; store on ice OR at -80°C. [Need to neutralize the MPA to pH 4~6, then analyze the undiluted sample with the kit]



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