Recombinant full length Human Calbindin protein with a His tag (His-Calbindin) purified from E. coli
IMR32 cell lysate.
This product was changed from ascites to tissue culture supernatant on 18th September 2017. Lot numbers higher than GR316455 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.
Shipped at 4°C. Store at 4°C prior to reconstitution. Store at -20°C. Stable for 12 months at -20°C.
Western blot - Calbindin antibody [AF2E5] (ab75524)
All lanes : Anti-Calbindin antibody [AF2E5] (ab75524) at 1/500 dilution
Lane 1 : Human kidney tissue lysate - total protein (ab30203) Lane 2 : Kidney (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 30 kDa Observed band size: 30 kDa
Exposure time: 90 seconds
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Calbindin antibody [AF2E5] (ab75524)This image is courtesy of an Abreview submitted by Karine Thibault
ab75524 staining rat brain (cortex) sections by IHC-FoFr. The animal was perfused with 4% paraformaldehyde and further post fixed with 4% paraformaldehyde overnight. The tissues were cryoprotected with 30% sucrose and sectioned using a cryostat. Staining with ab75524 at a 1/2800 dilution in PBS-Triton (0.3%) with 0.02% azide was performed for 18h at 25°C. A donkey anti-mouse Alexa488 polyclonal antibody at 1/1000 was used as the secondary antibody.
Overlay histogram showing SH-SY5Y cells stained with ab75524 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75524, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
ICC/IF image of ab75524 stained Hek293 cells. The cells were 4% PFA fixed (10mins) 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 (ab75524, 1:1000 dilution) overnight at +4°C. The secondary antibody (green) was ab96879 Dylight 488 goat anti-mouse IgG (H+L) used at a 1/250 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.