Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (AB222391)

IHC image of ab32535 staining in human breast cancer formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32535, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32535).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (AB222391)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Lamin B Receptor/LBR with unpurified ab32535 at 1/20 dilutionc (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32535).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (AB222391)

ab32535, at a 1/500 dilution, staining Lamin B Receptor/LBR in HeLa cells by Immunofluorescence.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32535).

• WB

Lab

Western blot - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (AB222391)

This data was developed using the same antibody clone in a different buffer formulation (ab32535).

Lanes 1 - 2 : Merged signal (red and green). Green - ab32535 observed at 58 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab32535 was shown to react with Lamin B Receptor/LBR in wild-type HEK-293 cells in western blot. Loss of signal was observed when LBR knockout cell lysate ab257503 was used. Wild-type and LBR knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk before incubation with ab32535 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Lamin B Receptor/LBR antibody [E398L] (<a href='/en-us/products/primary-antibodies/lamin-b-receptor-lbr-antibody-e398l-ab32535'>ab32535</a>) at 1/500 dilution

Lane 1:

Wild-type HEK-293 cell lysate at 20 µg

Lane 2:

Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-lbr-lamin-b-receptor-knockout-hek-293t-cell-lysate-ab257503'>ab257503</a>) at 20 µg

Lane 2:

Western blot - Human LBR (Lamin B Receptor) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-lbr-lamin-b-receptor-knockout-hek-293t-cell-line-ab266308'>ab266308</a>)

Predicted band size: 71 kDa

Observed band size: 58 kDa

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