This antibody gave a positive siganl in the following Human Lysates:
HCT 116 Whole Cell, Jurkat Whole Cell, HEK293 Whole Cell, HepG2 Whole Cell, Colon Tissue Lysate, Kidney Tissue Lysate, Liver Tissue Lysate
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
ICC/IF image of ab64071 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab64071, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293, and MCF-7 cells at 1µg/ml, and in 100% Methanol fixed (5 min) HeLa, Hek293, HepG2, and MCF-7 cells at 5µg/ml.
Western blot - Anti-HINT1 antibody (ab64071)
All lanes : Anti-HINT1 antibody (ab64071) at 1 µg/ml
Lane 1 : HCT 116 (Human Colorectal Carcinoma) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 5 : Human colon tissue lysate - total protein (ab30051) Lane 6 : Human kidney tissue lysate - total protein (ab30203) Lane 7 : Human liver tissue lysate - total protein (ab29889)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 14 kDa Observed band size: 14 kDa Additional bands at: 60 kDa, 70 kDa, 90 kDa. We are unsure as to the identity of these extra bands.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HINT1 antibody (ab64071)This image is courtesy of an abreview submitted by Aamir Ahmed, University College London, United Kingdom
IHC-P image of Hint1 (ab64071) on Colon cancer sections. The sections were fixed in paraformaldehyde and underwent heat mediated antigen retrieval using Novacastra (pH6). The sections were then blocked in 0.3% H2O2 solution for 10 mins at 22°C.