i want to data of MTT assay of HepG2 - deos my reading are relevant? please help ! (Nov/11/2006 )
hell o a ll,
i salute to the all the mentors of this bioforum who are always ready to help others.
i did the MTT assay on the HepG2 cells. and as i know now how to interpret the data.
i seeded diff cell no of HepG2/well .
after 24 hr , i changed the medium
next 24 hr i added 20 microlitr of the 5mg per ml MTT to each well.
then 2 hr after, i remove the MTT containing medium
wash wih PBS
then i add 100 micro liter DMSO tp each to dissolve the formazan crystal.
after overnite incubation, i got the read thru the ELISA reader at 540 nm.
my reading are aass following.
cell seeded per well reading
2500------------------------------.117
7500------------------------------0.261
10,000---------------------------0.267
15000---------------------------0.275
17,500--------------------------0.395
20,000--------------------------.443
now , i want to ask some thing about my reading that whther my reading are significant of not.
is there any scientist who is also workin on the HepG2 cell and and performing the MTT assay. i want to ask what is the duplication of correct time of the Hepg2 .
any fruitfull knowledge related my experiment is highly appreciated.
thanking you.
i salute to the all the mentors of this bioforum who are always ready to help others.
i did the MTT assay on the HepG2 cells. and as i know now how to interpret the data.
i seeded diff cell no of HepG2/well .
after 24 hr , i changed the medium
next 24 hr i added 20 microlitr of the 5mg per ml MTT to each well.
then 2 hr after, i remove the MTT containing medium
wash wih PBS
then i add 100 micro liter DMSO tp each to dissolve the formazan crystal.
after overnite incubation, i got the read thru the ELISA reader at 540 nm.
my reading are aass following.
cell seeded per well reading
2500------------------------------.117
7500------------------------------0.261
10,000---------------------------0.267
15000---------------------------0.275
17,500--------------------------0.395
20,000--------------------------.443
now , i want to ask some thing about my reading that whther my reading are significant of not.
is there any scientist who is also workin on the HepG2 cell and and performing the MTT assay. i want to ask what is the duplication of correct time of the Hepg2 .
any fruitfull knowledge related my experiment is highly appreciated.
thanking you.
ideal culture conditions promote doubling of HepG2 after ca. 29h;
what is missing is a background control which should subtracted from data values; a positive control to show that your test is working is also missing; to evaluate if there is significancy you have to perform the assays for each value at least three times, and the MTT test itself is to perform also at least three times to get n=3; then use standard statistics f.i. ANOVA if there are at least three groups