Anti-LAMP1 antibody [25/Lamp-1] - Lysosome Marker

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-LAMP1 antibody [25/Lamp-1] - Lysosome Marker (AB289548)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling LAMP1 with ab289548 at 1/1000 dilution (0.1 μg) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti-Mouse IgG (Alexa Fluor^® 488, ab150113) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-LAMP1 antibody [25/Lamp-1] - Lysosome Marker (AB289548)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labelling LAMP1 with ab289548 at 1/100 (10 μg/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image shows cytoplasmic staining in HeLa cell line. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/500 dilution (Red). The nuclear counterstain was DAPI (Blue).

Negative control 1 : ab289548 at 1/100 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/500 dilution.

Negative control 2 : ab179513 Anti-beta Tubulin rabbit monoclonal antibody at a 1/1000 dilution followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• WB

Lab

Western blot - Anti-LAMP1 antibody [25/Lamp-1] - Lysosome Marker (AB289548)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 27061067, 15111122).

Exposure time : 3 seconds

All lanes:

Western blot - Anti-LAMP1 antibody [25/Lamp-1] - Lysosome Marker (ab289548) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

LAMP1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 45 kDa

Observed band size: 110 kDa

false

• WB

Lab

Western blot - Anti-LAMP1 antibody [25/Lamp-1] - Lysosome Marker (AB289548)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 27061067, 15111122).

Exposure time : Lane 1, 4, 5 : 26 seconds ; Lane 2, 3 : 70 seconds

All lanes:

Western blot - Anti-LAMP1 antibody [25/Lamp-1] - Lysosome Marker (ab289548) at 1/1000 dilution

Lane 1:

Human kidney tissue lysate at 20 µg

Lane 2:

Human spleen tissue lysate at 20 µg

Lane 3:

Human colon cancer tissue lysate at 20 µg

Lane 4:

HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 20 µg

Lane 5:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 45 kDa

Observed band size: 110 kDa

false