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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling LAMP1 with ab278043 at 1/100 (5.45 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 (green). Confocal image showing cytoplasmic staining in HeLa. ab25630 Anti-LAMP1 mouse monoclonal antibody was used to counterstain at 1/500 (red). The Nuclear counterstain was DAPI (blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human thyroid tissue staining FUCA1 with ab323794 at a 1 : 1000 (0.511 ug/ml) dilution, ab278043 anti-LAMP1 used at 1 : 5000 (0.103 ug/ml) dilution.

Panel A : merged staining of anti-FUCA1 (green; Opal™520), anti-LAMP1 (magenta; Opal™570) on human thyroid.
Panel B : anti-FUCA1 showed granular staining in human thyroid.
Panel C : ant-LAMP1 staining lysosome in human thyroid.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323794 and ab278043 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labelling LAMP1 with ab278043 at 1/5000 (0.109 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Cytoplasmic staining in human lung cancer. The section was incubated with ab278043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling LAMP1 with ab278043 at 1/5000 (0.109 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Cytoplasmic staining in human liver. The section was incubated with ab278043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labelling LAMP1 with ab278043 at 1/5000 (0.109 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Cytoplasmic staining in human pancreas. The section was incubated with ab278043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

Immunohistochemical analysis of paraffin-embedded Human tonsil labelling LAMP1 with ab278043 at 1/5000 (0.109 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Cytoplasmic staining in human tonsil. The section was incubated with ab278043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

Intracellular flow cytometric analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labelling LAMP1 with ab278043 at 1/500 dilution (red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

Western blot : Rabbit monoclonal [EPR24395-31] to LAMP1 ab278043 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 80-120 kDa in Wild-type A549 and HCT 116 cell lysates with no signal observed at this size in LAMP1 knockout A549 cell line and LAMP1 knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

Lanes 1 - 4:

Western blot - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (<a href='/ko/products/primary-antibodies/lamp1-antibody-epr24395-31-bsa-and-azide-free-ab278046'>ab278046</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (ab278043) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

Western blot - Human LAMP1 knockout A549 cell line (<a href='/ko/products/cell-lines/human-lamp1-knockout-a549-cell-line-ab288919'>ab288919</a>) at 20 µg

Lane 3:

Wild-type HCT 116 ab277289 cell lysates at 20 µg

Lane 4:

Western blot - Human LAMP1 knockout HCT116 cell line (<a href='/ko/products/cell-lines/human-lamp1-knockout-hct116-cell-line-ab289151'>ab289151</a>) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 80-120 kDa

false

• WB

Lab

Western blot - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

Western blot : Anti-LAMP1 antibody [EPR24395-31] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab278043 was shown to bind specifically to LAMP1. A band was observed at 100-120 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in LAMP1 knockout cell line. To generate this image, wild-type and LAMP1 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (ab278043) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

LAMP1 knockout HCT 116 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Observed band size: 100-120 kDa

false

• WB

Lab

Western blot - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

Blocking and Diluting buffer : 5% NFDM/TBST

LAMP1 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 27061067, 15111122).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (ab278043) 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

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 45 kDa

Observed band size: 110 kDa

false

• WB

Lab

Western blot - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of HeLa whole cell lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab278043 (1/2000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG (H+L) (ab97051) at 1/50,000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique.

ab278043 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected under these conditions. This data was generated as part of a structured antibody stability study supporting Abcam’s transition from cold‑chain to ambient shipping.

All lanes:

Western blot - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (ab278043) at 1/2000 dilution

All lanes:

HeLa whole cell lysate at 15 µg with NDFM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

false

Exposure time: 40s

• WB

Lab

Western blot - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (AB278043)

Blocking and Diluting buffer : 5% NFDM/TBST

LAMP1 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 27061067, 15111122).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker (ab278043) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 45 kDa

Observed band size: 110 kDa

false