Anti-LMAN1 antibody [EPR6979]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-LMAN1 antibody [EPR6979] (AB125006)

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).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] (AB125006)

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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-LMAN1 antibody [EPR6979] (AB125006)

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] (AB125006)

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] (AB125006)

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

Lab

Western blot - Anti-LMAN1 antibody [EPR6979] (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] (ab125006) 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

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• WB

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Western blot - Anti-LMAN1 antibody [EPR6979] (AB125006)

All lanes:

Western blot - Anti-LMAN1 antibody [EPR6979] (ab125006) 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

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• WB

CiteAb

Western blot - Anti-LMAN1 antibody [EPR6979] (AB125006)

LMAN1 western blot using anti-LMAN1 antibody [EPR6979] ab125006. Publication image and figure legend from Emmer, B., Hesketh, G. G., et al., 2018, Elife, PubMed 30251625.

ab125006 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab125006 please see the product overview.

SURF4 localizes to the early secretory pathway where it physically interacts with PCSK9.(A) Immunofluorescence of FLAG-SURF4 together with markers of the ER (calnexin), ERGIC (LMAN1), and Golgi (GM130). Scale bar = 10 µm. (B) Quantification of colocalization (n = 8 cells analyzed for each combination of antibody staining). (C) Spectral counts for SURF4 in streptavidin-purified eluates from cells expressing various BirA* fusion proteins. (D) Immunoprecipitations were performed using antibodies directed against FLAG or GFP from lysates of cells expressing FLAG-SURF4, PCSK9-eGFP, both, or neither. Error bars represent standard deviations.

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