Jackson ImmunoResearch
Alexa Fluor® 488 AffiniPure Donkey Anti-Mouse IgG (H+L) (min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Rb, Rat, Shp Sr Prot)
715-545-151
Polyclonal
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I coated cover glasses with rat anti-MHCII antibodies and seeded dendritic cells on them to see the surface expression of several receptors within the engagement of MHCII by immunostaining. Unfortunately, I used this secondary antibody, sold to be minimal crosslinking with Rat, and it WAS CROSS-REACTING with my rat coating. I lost time and effort trying to work with this antibody, but nothing worked. I strongly do not recommend this antibody if you have to work with rat proteins.
Immunofluorescence
Bone marrow derived dendritic cells (mouse)
None
BSA 5% tested (1h at RT)
Phalloidin 647 1/250 + Donkey Anti-Mouse IgG (H+L)1/250 (1h at RT) 488
TIRF
Cross-linking of the secondary antibody onto the rat-antibody coated cover glass. I also tested to block with Donkey serum, even overnight I have got the same results. I used a fibronectin coating and an Armenian hamster antibody coating as controls. There was no sign of a cross-reaction then.
N/A
Good fluorescence.
Not specific.
Do not use it with Rat proteins.