PE Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - PE Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB281778)

Flow cytometry overlay histogram showing wild-type Hap1 (green line) and LAMP1 knockout Hap1 cells stained with ab281778 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab281778) (1x 10⁶ in 100μl at 0.04μg/ml (1/12500) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody (wild-type Hap1 - black line LAMP1 knockout Hap1- grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5000 events were collected using a 50 mW Yellow/Green laser (561nm) and 585/42 bandpass filter.

This antibody can also be used in Hap1 cells fixed with 4% formaldehyde (10 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - PE Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB281778)

ab281778 staining LAMP1 in HAP1 cells, with negative expression in LAMP1 knockout HAP1 cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab281778 at 0.2 μg/ml (shown in green). ab190573, rabbit monoclonal to alpha Tubulin (Alexa Fluor^® 647) (shown in magenta) was used at 5 μg/ml for structural counterstaining.

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.