Anti-LAMP2 antibody [EPR19512] - Lysosome Marker

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (AB199946)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling LAMP2 with ab199946 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human kidney is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (AB199946)

Immunohistochemical analysis of paraffin-embedded Human thyroid cancer tissue labeling LAMP2 with ab199946 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human thyroid cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (AB199946)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 and 2 : 15 seconds; Lane 3,4 and 5 : 5 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID : 19828315) and lots of other products.

All lanes:

Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946) at 1/2000 dilution

Lane 1:

Human fetal liver lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Lane 4:

Human fetal spleen lysate at 10 µg

Lane 5:

Human placenta lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 45 kDa

Observed band size: 110-130 kDa

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• WB

Unknown

Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (AB199946)

Lanes 1 - 4 : Merged signal (red and green). Green - ab199946 observed at 110 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab199946 was shown to react with LAMP2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255402 (knockout cell lysate ab263861) was used. Wild-type and LAMP2 knockout samples were subjected to SDS-PAGE. ab199946 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 μg/ml and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946) at 1 µg/mL

Lane 1:

Wild-type HEK-293 cell lysate at 20 µg

Lane 2:

LAMP2 knockout HEK-293 cell lysate at 20 µg

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

LAMP2 knockout HeLa cell lysate at 20 µg

Predicted band size: 45 kDa

Observed band size: 110 kDa,37 kDa

false

• WB

Lab

Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (AB199946)

Lanes 1 - 4 : Merged signal (red and green). Green - ab199946 observed at 45 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab199946 was shown to recognize LAMP2 in wild-type HEK-293 cells as signal was lost at the expected MW in LAMP2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and LAMP2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab199946 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946) at 1/2000 dilution

Lane 1:

Wild-type HEK-293 whole cell lysate at 20 µg

Lane 2:

LAMP2 knockout HEK-293 whole cell lysate at 20 µg

Lane 3:

HepG2 whole cell lysate at 20 µg

Lane 4:

THP1 whole cell lysate at 20 µg

Predicted band size: 45 kDa

false

• WB

Supplier Data

Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (AB199946)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1,2 and 3 : 1 second; Lane 4 : 5 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID : 19828315) and lots of other products.

All lanes:

Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946) at 1/2000 dilution

Lane 1:

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

Lane 2:

THP-1 (Human monocytic leukemia cell line) whole cell lysate at 20 µg

Lane 3:

JAR (Human placenta choriocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cell line from peripheral blood) 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 1/100000 dilution

Predicted band size: 45 kDa

Observed band size: 110-130 kDa

false