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Janus Green cell normalization stain (ab111622)

Submit a review Q&A (2)References (1)

Overview

  • Product name

    Janus Green cell normalization stain

  • Tested applications

    Suitable for: In-Cell ELISAmore details

  • General notes

    Ready-to-use green stain for normalizing antibody signal intensity to total cell amounts during in-cell ELISA (ab111622).

    Cells fixed to a solid surface (typically in microplate wells) can be stained by adding 1X Janus Green Stain. Incubate cells for 5 minutes at room temperature. Dye should be removed and washed 5 times in ultrapure water or until excess dye is removed. The last wash should be removed and add 0.1 mL 0.5 M HCl per well and incubate for 10 minutes. Shake the plate for 10 seconds, and measure the OD 595 nm using a standard microplate spectrophotometer.

Properties

  • Form

    Liquid

  • Storage instructions

    Store at +4°C.

  • Storage buffer

    Constituents: 0.02% Monobasic dihydrogen potassium phosphate, 0.216% Dibasic monohydrogen sodium phosphate, 0.02% Potassium chloride, 0.81% Sodium chloride, Water, 0.3% Janus Green B
  • Concentration information loading...

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab111622 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
In-Cell ELISA
Use at an assay dependent dilution. 50 µL/ well fixed cells
Notes
In-Cell ELISA
Use at an assay dependent dilution. 50 µL/ well fixed cells

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (1)

Publishing research using ab111622? Please let us know so that we can cite the reference in this datasheet.

ab111622 has been referenced in 1 publication.

  • Liang KX  et al. Disease-specific phenotypes in iPSC-derived neural stem cells with POLG mutations. EMBO Mol Med 12:e12146 (2020). PubMed: 32840960

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Question

For this janus green assay, the protocol states to read the plate at 595nM. Can the absorbance of the 0.1 ml HCl solution be read in the same wells the cells were grown or do you recommend spinning down the plate, removing the solution to clean well prior to taking OD measurements?

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Answer

Yes, the OD 595 can be read directly on the plate where the cells were grown and the assay was performed. The HCL will disrupt the cells and the janus green (which stains the nuclei) will be released into the well. No need to spin or move the HCL to a different plate.

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Question

Dear Sirs,
a customer of ours would like to know which is the right wavelenght to read the signal of Janus Green.

Thanks in advance
Kind regards

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Answer

Thank you for your enquiry,

We recommend to measure the assays results at OD595 nm using a standard microplate spectrophotometer. We will be amending the datasheet to make this clearer.

Whole Cell Staining with Janus Green:

1. Completely remove HRP Development Solution from themicroplate wells and blot to dry. Add 50 μl of 1X JanusGreen Stain per well. Incubate plate for 5 minutes atroom temperature.

2. Remove dye, wash plate 5 times in ultrapure water oruntil excess dye is removed.

3. Remove last water wash, blot to dry, add 100 μl of 0.5 MHCl and incubate for 10 minutes.

4. Shake the plate for 10 seconds, and measure the OD595 nm using a standard microplate spectrophotometer

I hope this will be helpful. If you have any further questions, please do not hesitate to contact us.

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