Anti-KIF1A antibody [EPR11790(B)] (ab180153)

Blocking and diluting buffer and concentration: 5% NFDM /TBST.

Immunocytochemistry/Immunofluorescence analysis of U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling KIF1A with purified ab180153 at 1/100 dilution (9.2μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilised with 0.1% tritonX-100. Ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used as the secondary antibody at 1/200 dilution (2.5 μg/ml). DAPI was used as the nuclear counter stain.

ab180153 (purified) at 1/50 dilution (2ug) immunoprecipitating KIF1A in SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate.

Lane 1 (input): SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate 10ug

Lane 2 (+): ab180153 + SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate 10ug

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab180153 in SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

For western blotting, ab131366 VeriBlot for IP secondary antibody (HRP) was used as the secondary antibody (1/10000). 

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling KIF1A with purified ab180153 at 1/90 dilution (10 ug/ml). Cells were fixed with 4% paraformaldehyde. A goat anti-rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/2000 dilution. Black - Isotype control, Rabbit monoclonal IgG. Blue - unlabelled control, cells without incubation with primary antibody and secondary antibody.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Immunofluorescent analysis of 4% paraformaldehyde-fixed 293 cells labeling KIF1A with unpurified ab180153 at 1/100 dilution, followed by Goat anti rabbit IgG (Dylight 555) at 1/250 dilution. Counter stained with Dapi.

Western blot analysis of 293 cell lysate immunoprecipitated using unpurified ab180153 at 1/30 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution.

Flow cytometric analysis of 2% paraformaldehyde-fixed SH-SY5Y cells labeling KIF1A with unpurified ab180153 at 1/150 dilution (red), compared to a Rabbit IgG control (green), followed by Goat anti rabbit IgG (FITC) at 1/75 dilution.