This antibody gave a positive result in the following whole cell lysates:
Jurkat (Human T cell lymphoblast-like cell line)
A431 (Human epithelial carcinoma cell line)
HEK293 (Human embryonic kidney cell line)
MCF7 (Human breast adenocarcinoma cell line)
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
ab24170 staining Lamp-1 in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab24170 at 1μg/ml and ab7291 (staining Tubulin) at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI, which was added to the secondary antibody mixture.
NOTE: The filamentous pattern observed only occurs when cell are fixed with 100% methanol. We are aware that the staining pattern can change if the cells are fixed with 4%PFA or an alternative fixative. LAMP-1 is a lysosome marker but also plays a role in cellular adhesion. Therefore, lysosomal and filamentous patterns for LAMP-1 can be possible.
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 - Lysosome Marker (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)This image is courtesy of an Abreview submitted by Mr Carl Hobbs
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.
Immunocytochemistry/ Immunofluorescence - LAMP1 antibody (ab24170)This image is courtesy of an anonymous abreview.
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).
Immunocytochemistry/ Immunofluorescence - Anti-LAMP1 antibody - Lysosome Marker (ab24170)This image is courtesy of an anonymous Abreview.
ab24170 staining LAMP1 in Human Retinal Microvascular Endothelial Cells (HRMEC) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton in PBS and blocked with 10% serum for 30 minutes at 22°C. Samples were incubated with primary antibody (1/300 in PBS) for 1 hour at 22°C. A diluted (1/500) Alexa Fluor® 488-conjugated Donkey anti-rabbit IgG polyclonal was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-LAMP1 antibody - Lysosome Marker (ab24170)Image is courtesy of an AbReview submitted by Veronica Winkelmann.
Immunocytochemical immunofluorescence analyis of PFA-fixed rat primary alveolar type II cells, labelling LAMP1 with ab24170 at a dilution of 1/200 incubated for 1/200. Blocking was with 10% BSA incubated for 5 minutes. Secondary was a donkey anti-rabbit polyclonal Alexa Fluor® 647 at 1/400.
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 +4°C. 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody (ab24170)Image courtsey Dr. Martin Broadstock (King's College London, Unted Kingdom)
IHC-P image of LAMP1 staining on human Cortex sections using ab24170 (1:400). The sections were deparaffinized and subjected to heat mediated antigen retreival using citric acid. The sections were then permeabilized using 0.05% Tween-20 and blocking was performed using 3% BSA for 1 hour at 21°C. The primary antibody ab24170 was diluted using 3% BSA with 0.05% Tween-20 in PBS and incubated with the sections for 18 hours at 4°C. The secondary antibody used was Goat polyclonal to rabbit IgG conjugated to biotin (1:500)