Cell Signalling
IL-6 (D3K2N) Rabbit antibody
12153
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Present study was designed to test the effect of different treatment on tissue specific IL-6 levels in diabetic rats. Proteins from rat aorta, liver, muscle, heart, Kidney and retina were extracted and an equal amount of proteins were resolved in 10% SDS-PAGE. Resolved proteins were immunoblotted for IL-6 and after anti species HRP-conjugated secondary antibody incubation, blots were developed using enhanced chemiluminescent reagent. Although, no species reactivity of antibody with rat was mentioned in antibody flyer, based on previous experience with cell signaling technology antibody was selected.
Western Blot
Rat tissue protein lysate
Overnight at 4 0C (5% Skimmed milk in TBS with 0.1% Tween 20; dilution 1:1000)
5 % Skimmed milk in TBS with 0.1% Tween 20 (for 1hr at RT)
2hr at RT (5% skimmed milk in TBS with 0.1% Tween 20; dilution 1:4000)
None
ECL
Although, antibody flyer does not mention antibody reactivity with rat proteins, strong, very specific and bright bands of IL-6 were observed at nearly 24KDa. This indicated that antibody is also reactive to rat proteins. Few non specific bands at higher molecular weight were also observed.
Gave highly specific band of IL-6 at correct molecular weight
Presence of some non specific band
Highly specific of Cell Signalling antibody for IL-6