Immunocytochemistry/ Immunofluorescence - LAMP1 antibody (ab24170)

ICC/IF image of ab24170 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab24170, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).

Western blot - LAMP1 antibody (ab24170)

All lanes : Anti-LAMP1 antibody (ab24170) at 1 µg/ml

Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed at 1/3000 dilution

Performed under reducing conditions.

Predicted band size : 120 kDa
Observed band size : 120 kDa
Additional bands at : 23 kDa,35 kDa,45 kDa. We are unsure as to the identity of these extra bands.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - LAMP1 antibody (ab24170)

ab24170 staining LAMP1 in human kidney tissue sections. Staining correlates with lysosomal specificity, particularly in the proximal convoluted tubules where lysosomes are enriched. Formalin/PFA-fixed human kidney tissue sections were incubated with ab24170 (1/200) for 2 hours. Antigen retrieval was performed by heat induction in citrate buffer pH 6. Please see accompanying abreview for additional information.

This image is courtesy of an Abreview submitted by Mr Carl Hobbs

Immunocytochemistry/ Immunofluorescence - LAMP1 antibody (ab24170)

ab24170 staining LAMP1 in human fibrosarcoma cells by Immunocytochemistry/ Immunofluorescence. The cells were PFA fixed, permeabilised in 0.1% Triton X-100 and incubated with the primary antibody at 1µg/ml for 1 hour at 20°C. The secondary antibody used was a donkey anti-rabbit (H+L) IgG conjugated to Alexa Fluor® 555 used at a 1/500 dilution. DAPI was used to stain the cell nuclei (blue).

This image is courtesy of an anonymous abreview.

Western blot - LAMP1 antibody (ab24170)

Anti-LAMP1 antibody (ab24170) at 1/700 dilution (in 5% milk for 4 hours at 20°C) + Rat Kidney - whole tissue lysate at 18 µg

Secondary
An HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 120 kDa
Observed band size : 120 kDa


Exposure time : 5 minutes

Blocking Step: 5% Milk for 1 hour at 20°C

This image is courtesy of an anonymous Abreview

Immunocytochemistry/ Immunofluorescence - Anti-LAMP1 antibody (ab24170)

ICC/IF image of ab24170 stained HeLa cells. The cells were 4% formaldehyde 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 (ab24170, 1µg/ml) overnight at +4ºC. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit 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.