Anti-LMAN1 antibody [EPR6979] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-LMAN1 antibody [EPR6979] - BSA and Azide free (AB248096)

This data was developed using ab125006, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling LMAN1 with Purified ab125006 at 1/20 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-LMAN1 antibody [EPR6979] - BSA and Azide free (AB248096)

This data was developed using ab125006, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of 293T (Human embryonic kidney epithelial cell) cells labeling LMAN1 with Purified ab125006 at 1/50 dilution (2 µg/mL). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] - BSA and Azide free (AB248096)

This data was developed using ab125006, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling LMAN1 with Purified ab125006 at 1/500 dilution (0.21 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] - BSA and Azide free (AB248096)

This data was developed using ab125006, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling LMAN1 with Purified ab125006 at 1/500 dilution (0.21 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] - BSA and Azide free (AB248096)

This data was developed using ab125006, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling LMAN1 with Purified ab125006 at 1/500 dilution (0.21 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.

• WB

Unknown

Western blot - Anti-LMAN1 antibody [EPR6979] - BSA and Azide free (AB248096)

All lanes:

Western blot - Anti-LMAN1 antibody [EPR6979] (<a href='/en-us/products/primary-antibodies/lman1-antibody-epr6979-ab125006'>ab125006</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

Mouse spleen lysate at 20 µg

Lane 4:

Mouse heart lysate at 20 µg

Lane 5:

Rat spleen lysate at 20 µg

Lane 6:

Rat heart lysate at 20 µg

Lane 7:

HEK-293 (Human embryonic kidney 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: 58 kDa

false

• WB

Lab

Western blot - Anti-LMAN1 antibody [EPR6979] - BSA and Azide free (AB248096)

This data was developed using the same antibody clone in a different buffer formulation (ab125006).

Lanes 1-4 : Merged signal (red and green). Green - ab125006 observed at 55 kDa. Red - loading control ab8245 observed at 37 kDa.

ab125006 Anti-LMAN1 antibody [EPR6979] was shown to specifically react with Protein ERGIC-53 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266248 (knockout cell lysate ab257505) was used. Wild-type and Protein ERGIC-53 knockout samples were subjected to SDS-PAGE. ab125006 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°CC at 1 in 1000 Dilution 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-LMAN1 antibody [EPR6979] (<a href='/en-us/products/primary-antibodies/lman1-antibody-epr6979-ab125006'>ab125006</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

LMAN1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human LMAN1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-lman1-knockout-hek-293t-cell-line-ab266248'>ab266248</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 58 kDa

Observed band size: 55 kDa

false