Application
Western blot
Sample
Rat Cell lysate - whole cell (Rat Trophoblast stem cell and human trophoblast ce)
Loading amount
100 µg
Specification
Rat Trophoblast stem cell and human trophoblast ce
Treatment
0.5% oxygen for 24 hours
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Concentration
1 µg/ml
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: Milk
Secondary antibody
Name
Non-Abcam antibody was used: anti rabbit
Host species: Goat
Clonality: Polyclonal
Conjugation: HRP polymer
Host species: Goat
Clonality: Polyclonal
Conjugation: HRP polymer
Dilution
1/5000
Detection
Detection method
ECL
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 171 kDa Non-specific: some kDa
Positive control
Human lysates at similar low oxygen concentrations.
Additional data
Additional Notes
This antibody works as predicted in human cell lysates, however it does not recognise rat protein (Western blot enclosed).
Abcam response
Thank you for submitting your review. Your data is quite interesting. You clearly show little to no KDM3A / JHDM2A is detected in the rat trophoblast stem cells. However, the human immunogen sequence used to generate this polyclonal antibody is 98% identical to the corresponding region in rat protein. Thus in theory ab80598 should recognize rat KDM3A / JHDM2A. Perhaps this antibody does not recognize rat KDM3A / JHDM2A or, possibly, expression levels of KDM3A / JHDM2A in this cell type are so low as to be undetectable by WB.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
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投稿 Jan 27 2011