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Our Abpromise guarantee covers the use of ab95470 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 0.3 - 1 µg/ml. Detects a band of approximately 28 kDa (predicted molecular weight: 25 kDa).|
ab95470 stained RAW 246.7 cells. The cells were 100% methanol fixed for 5 minutes at-20°C and then incubated in 1%BSA / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab95470 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) preadsorbed (ab150133) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Immunocytochemical immunoflurescence analysis of PFA-fixed human neuroblastoma cell line, labelling TREM2 using ab95470 at a dilution of 1/200 incubated for 2 hours and 24°C in 0.1M PBST with 10% donkey serum. Permeablisation was accomplished using 0.1M PBS with 1% Triton X. Blocking was done with 10% donkey serum in PBST and incubated for 30 minutes at 24°C. The secondary used was a polyclonal donkey anti-goat Alexa Fluor® 568 conjugate.