Here is a SDS PAGE of Jurkat total lysate. Can anyone suggest what is the highly expressed protein? Unfortunately a marker is absent.
The highly expressed protein could actually be albumin from the cell media. Most media preparations include serum, and albumin is an abundant serum protein.
I might be misunderstanding the question.
That's an a protein being overexpressed at levels some people would only dream of. I might be way off here, but I do not believe that those are unmodified Jurkat cells. See, for comparison, http://ow.ly/olthK where the Jurkat whole cell lysate is lane number 2.
So if you meant to imply that those are unmodified cells growing under standard conditions, you might want to consider that those cells are misidentified.
HTH,
August
The highly expressed protein could actually be albumin from the cell media. Most media preparations include serum, and albumin is an abundant serum protein.
Just by looking at the gel, you can not say which protein it is. Have you tried to cut it off and examine it by LC-MS/MS?
Cells were washed with PBS, so it shouldn't be an albumin from medium.
I haven't done MS of this thing yet. I was also surprised when I saw such a band, so I thought maybe Jurkat realy overexpress something.
Maybe this band corresponds to three distinct bands between 35 and 55 kDa on August Pamplona's gel, if they were not separated enough..
If there's any doubt about the washing procedure, replicate the procedure with a distinct but similarly cultured cell line (if available) and see what the gel shows then. If nothing else, continuing to find the band in the apparent Jurkat line but not in the comparison cell line should rule out a medium contaminant as suggested by Anne.
P.S. Not my gel. Just a random gel from a supply house with a Jurkat lane.
Hi , Thanks for sharing this problem
Mine is similar problem i am attaching gel which shows Jurkat cell lysate in Ponceau s where you can see thick blobs. Whenever i do western blot my antibody picks this blob and unfortunately my protein fall in that region. i tried washing and lysing in different buffers but though the blob intensity decreases my western always picks up that blob. please help if anybody could solve this problem.
I made phoresis of RPMI medium. Yes, this band comes from the medium. Cells should have been washed more properly.. Thank you for solving the problem.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology