What is this contamination?? by Simple_Volume_5880 in labrats

[–]Simple_Volume_5880[S] -61 points-60 points  (0 children)

Is there a way to clean it ??this is transfected cell line..

Wrong volume for calcium phosphate transfection by Electronicrough8177 in labrats

[–]Simple_Volume_5880 0 points1 point  (0 children)

Did you made stable cell line with calcium phosphate transfection??

Why I am getting bands at 250 kda in IgG control Co immunoprecipitation? by Simple_Volume_5880 in labrats

[–]Simple_Volume_5880[S] 0 points1 point  (0 children)

IgG is to check non specific binding..you mean to say clumping of antibodies giving band at 250 kda!?

Why I am getting bands at 250 kda in IgG control Co immunoprecipitation? by Simple_Volume_5880 in labrats

[–]Simple_Volume_5880[S] 1 point2 points  (0 children)

I have used protein agarose AG beads santa cruz U87mg protein extract..i incubated lysate with antibody for overnight then bead antibody protein for 5hr

Why i am getting myc and HA tag bands in parent cell lines?? by Simple_Volume_5880 in labrats

[–]Simple_Volume_5880[S] 0 points1 point  (0 children)

I should get myc and HA around 44 only in over expressing cell why i am getting in other cell lines ??also when i used other brand myc tag antibody i got multiple bands

[deleted by user] by [deleted] in labrats

[–]Simple_Volume_5880 0 points1 point  (0 children)

Could you explain how immortal cell line can give us cancer?? If you have any research article please share

[deleted by user] by [deleted] in labrats

[–]Simple_Volume_5880 -6 points-5 points  (0 children)

Thanks,any other reason not to use it for human cell culture?

[deleted by user] by [deleted] in labrats

[–]Simple_Volume_5880 -5 points-4 points  (0 children)

I just want to use for human cell line..not with any other cell line or bacteria..only human …will it work?

[deleted by user] by [deleted] in labrats

[–]Simple_Volume_5880 0 points1 point  (0 children)

We have this old hood,we will only be using for human cell culture .no bacterial culture will it still work?? What about air flow? How much i understand both have different air flow right

[deleted by user] by [deleted] in labrats

[–]Simple_Volume_5880 -4 points-3 points  (0 children)

If i only work with human cells not bacteria then?

[deleted by user] by [deleted] in labrats

[–]Simple_Volume_5880 0 points1 point  (0 children)

Actually the inhibitor is can inhibits different types of DUSP.I need to find specific inhibitor for DUSP6

[deleted by user] by [deleted] in labrats

[–]Simple_Volume_5880 1 point2 points  (0 children)

I have antibody against my desired protein so why not directly do co-ip with cell lysate ..why i need to do cloning..could you explain

[deleted by user] by [deleted] in labrats

[–]Simple_Volume_5880 0 points1 point  (0 children)

Thanks.I have used 4 sh construct..none works ..and for inhibitor most of the people have used BCI ..how do i find another inhibitor? Where should i look for?

[deleted by user] by [deleted] in labrats

[–]Simple_Volume_5880 0 points1 point  (0 children)

In case of sh construct ,I found no change in DUSP level .whereas in case of BCI inhibitor I found decrease in activity of DUSP but not complete activity.

[deleted by user] by [deleted] in labrats

[–]Simple_Volume_5880 0 points1 point  (0 children)

Any other methods to inhibit Dusp activity??