Abcam
Anti-VEGFC antibody
ab9546
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In the present experiment, diabetic rats kept in different nutritional condition were subjected to hind limb ischemia by ligating femoral artery and cutting of distal to the inguinal ligament and proximal to the popliteal bifurcation site. After surgery, rats were sacrificed at different time points, femoral muscles were harvested, proteins were extracted, and an equal amount of proteins were resolved in 12% SDS-PAGE. Resolved proteins were western transferred in nitrocellulose membrane and probed for VEGFC levels. Selection of antibody from abcam was based on previous experience with same antibody.
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 2 hr at RT)
1hr at RT (5% skimmed milk in TBS with 0.1% Tween 20; dilution 1:5000)
None
ECL
Although, specific band of VEGFC were detected at nearly 22 KDa, lot of non-specific bands with bright intensities were also observed. Blot was not very clean as lots of other bands were also visible.
Got specific band of VEGFC in rat proteins
Lot of non-specificity detected
Average reactivity of Abcam VEGFC antibody to rat proteins