Hello,
I am always grateful for those who answer my questions.
I learn a lot thanks to such help.
I want to ask question about developer and fixer, required for western blot.
I asked quite a similar question indeed but i want to get some additional information.
From previous answeres, I learned quite well that some water
(Not much amount because the water is for washing.)
remained in containers of developer and fixer are not critical for
detection results in western blot.
But i guess that remnants in the containers of developer and fixer
are not only waters, but other things as well.(Etoh, tiny particles, and others.)
I wonder whether such things can affect detection results in western blot
in spite of little amount.
So far, I found no problems considering my detection results but
I want to make sure about what I have asked above.
Thank you!
Dear Lee
i think I provided an answer to a similar question a few months back. To reiterate my previous answer and frame in the context of your new query
- No. Providing you wash out your measuring cyclinders that have been used to dilute fixative and developer with fresh tap water residue in such cyclinders will not affect sensitivity of detection or background
- indeed with old style dark rooms and manual development in trays it is common for both the trays and measuring cyclinders to become discoloured ( brown) over time
- Do however separate your measuring cylinders for developer and fix and make sure you wash them out along with developing trays with fresh tap water EVERY TIME you use; both before and after use
- In terms is sensitivity the important thing is to shield your developer from the light to prevent photo oxidation: if your developer turns from translucent light brown towards opaque dark brown discard down sink: darker oxidised developer will reduce your sensitivity and increase background
- As mentioned in my previous answer it is ok to keep fixer and developer IN THE DARK for prolonged periods during the day but do not use the next day as this will increase chances of reduced sensitivity and increased bacground
- Residue in measuring cylinders will not have an effect unless it leaches or flakes into your diluted solutions. If you rinse just before use that won't happen
Dear Lee
i think I provided an answer to a similar question a few months back. To reiterate my previous answer and frame in the context of your new query
- No. Providing you wash out your measuring cyclinders that have been used to dilute fixative and developer with fresh tap water residue in such cyclinders will not affect sensitivity of detection or background
- indeed with old style dark rooms and manual development in trays it is common for both the trays and measuring cyclinders to become discoloured ( brown) over time
- Do however separate your measuring cylinders for developer and fix and make sure you wash them out along with developing trays with fresh tap water EVERY TIME you use; both before and after use
- In terms is sensitivity the important thing is to shield your developer from the light to prevent photo oxidation: if your developer turns from translucent light brown towards opaque dark brown discard down sink: darker oxidised developer will reduce your sensitivity and increase background
- As mentioned in my previous answer it is ok to keep fixer and developer IN THE DARK for prolonged periods during the day but do not use the next day as this will increase chances of reduced sensitivity and increased bacground
- Residue in measuring cylinders will not have an effect unless it leaches or flakes into your diluted solutions. If you rinse just before use that won't happen
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