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Protein Extraction from different cells organs

Protein Extraction from different cells organs - Western Blot Forum

Protein Extraction from different cells organs - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 02-13-2012, 02:18 AM
Pipette Filler
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Default Protein Extraction from different cells organs



Hi all!

I tried to detect protein with western blotting. previously, my colleague did the experiment and it worked. but when I applied the procedure to different cell organs, the result was not good, eventhough it was detectable.

despite my low skills as consideration, I wonder does cells/organ type give credits to the results (high backgroud, unspecific band, etc)?

specifically, is there any specific consideration for protein extraction from liver organs?

Thank you for your suggestion

Best regards
Rizky
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Old 02-15-2012, 04:31 AM
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Default Re: Protein Extraction from different cells organs

Hi Rizky, every tissue and organ is usually prepared, extracted and isolated differently as you know.

Organs especially are notoriously extremely difficult to extract proteins, as they require perfusion with enzymes to allow proper release/breakdown of the proteins from the cells.

Can you use a cell line or purchased lysate instead?

Also of note the liver contains a high amount of lipids in the form of VLDL/LDL and cholesterol/lipids and these may be interfering with your experiment at various phases.

It may be worthwhile to try ultracentrifugation or at least high RPM centrifugation to get most of the lipids out.
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Old 02-15-2012, 07:22 AM
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Default Re: Protein Extraction from different cells organs

Thank you for your response.

I used the tissue from experimental model. I think I couldnt use from cell line.

lipid is also my consideration. from Western blotting result, what kind of description that might be resulted from lipid interference?

Thank you
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Old 02-15-2012, 07:55 AM
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Default Re: Protein Extraction from different cells organs

Hi there, it depends on several factors:

1) is lipid affecting your concentration/isolation of the protein
2) is lipid affecting your experimental detection

Also of note what is the quality of protein extracted, purity and concentration.

Is your protein expressed highly in liver?, if so you should easily get a high signal if you can get high concentration of your protein and have accurate detection methods.
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Old 02-23-2012, 12:32 PM
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Default Re: Protein Extraction from different cells organs

probably it is better for me to attach the result. I will try to upload the pic

thank you

discussion continued here:

[Only registered users see links. ]

Last edited by admin; 02-24-2012 at 03:50 AM.
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Old 02-24-2012, 03:53 AM
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Default Re: Protein Extraction from different cells organs

Move western blot discussion here:

Western Blotting from Protein Extracted from Organs Tissue with High Lipid Content
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