Anti-LAMP1 antibody [EPR4204] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR4204] - BSA and Azide free (AB247693)

This data was developed using ab108597, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of LAMP1 in paraffin embedded Human kidney tissue, using unpurified ab108597 at a dilution of 1/100.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR4204] - BSA and Azide free (AB247693)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver carcinoma tissue sections labeling LAMP1 with purified ab108597 at 1/400 dilution (0.36 μg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108597).

• WB

Lab

Western blot - Anti-LAMP1 antibody [EPR4204] - BSA and Azide free (AB247693)

This data was developed using ab108597, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-LAMP1 antibody [EPR4204] - BSA and Azide free (ab247693) at 1/5000 dilution

Lane 1:

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

Lane 2:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

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='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 45 kDa

false

Exposure time: 5s

• WB

Unknown

Western blot - Anti-LAMP1 antibody [EPR4204] - BSA and Azide free (AB247693)

This data was developed using ab108597, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-LAMP1 antibody [EPR4204] - BSA and Azide free (ab247693)

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

JAR cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

A431 cell lysate at 10 µg

Predicted band size: 45 kDa

false

• WB

Supplier Data

Western blot - Anti-LAMP1 antibody [EPR4204] - BSA and Azide free (AB247693)

This data was developed using ab108597, the same antibody clone in a different buffer formulation.

Lanes 1 - 4 : Merged signal (red and green). Green - unpurified ab108597 observed at 110 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab108597 was shown to recognize LAMP1 in wild-type HAP1 cells as signal was lost at the expected MW in LAMP1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and LAMP1 knockout samples were subjected to SDS-PAGE. ab108597 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 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 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-LAMP1 antibody [EPR4204] - BSA and Azide free (ab247693) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

LAMP1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

MCF7 whole cell lysate at 20 µg

Lane 4:

Human Kidney whole cell lysate at 20 µg

Predicted band size: 45 kDa

Observed band size: 110 kDa

false