Immunocytochemistry/ Immunofluorescence - LAMP1 antibody [1D4B] - BSA and Azide free (ab25245)

ab25245 staining mouse macrophage, hepatoma cell by ICC/IF.  Cells were PFA fixed and permeabilized in Triton X-100 prior to blocking in 1% BSA for 30 minutes at 25°C.  The primary antibody was diluted 1/1000 and incubated with the sample for 16 hours at 4°C.  An Alexa Fluor® 594 conjugated goat anti-mouse antibody was used as the secondary.

This image is courtesy of an Abreview submitted by Dr Tian Xia

Immunocytochemistry/ Immunofluorescence - LAMP1 antibody [1D4B] - BSA and Azide free (ab25245)

ab25245 staining LAMP1 in Mouse epithelial cells by ICC/IF (Immunocytochemistry/immunoflurescence). Cells were fixed with methanol/acetone 1:1 and blocked with 1% milk in PNB for 1 hour at 25°C. Samples were incubated with primary antibody (1/500) in PNB for 16 hours at 4°C. An Alexa Fluor^® 568-conjugated Donkey polyclonal to rat IgG, dilution 1/500, was used as secondary antibody. Nuclei were stained blue.

This image is courtesy of an anonymous Abreview

Flow Cytometry-LAMP1 antibody [1D4B] - BSA and Azide free(ab25245)

Overlay histogram showing Jurkat cells stained with ab25245 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab25245, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.