Anti-LOXL2 antibody [7HCLC]

• ICC

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Immunocytochemistry - Anti-LOXL2 antibody [7HCLC] (AB277807)

For immunofluorescence analysis, HeLa cells were fixed and permeabilized for detection of endogenous LOXL2 using Anti-LOXL2 Recombinant Rabbit Polyclonal Antibody (ab277807, 5 μg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal^™ Secondary Antibody, Alexa Fluor^® 488 conjugate (1/2000). Nuclei (blue) is stained using SlowFade^® Gold Antifade Mountant with DAPI and cytoskeletal F-actin (red) staining using Rhodamine Phalloidin (1/300) Panel a-d) shows representative un-treated cells that were stained for detection and localization of LOXL2 protein (green) with less signal. Panel e-h) clearly demonstrate enhanced perinuclear localisation of LOXL2 in cells treated with Cobalt chloride (100M 16h). The images were captured at 60X magnification.

• WB

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Western blot - Anti-LOXL2 antibody [7HCLC] (AB277807)

Western blot analysis was performed on whole cell extracts (30 μg lysate) of HeLa (human epithelial cell line from cervix adenocarcinoma) (Lane 1), HeLa treated with cobalt chloride (100 M for 16 hrs) (Lane 2), U-87 MG (human glioblastoma-astrocytoma epithelial cell line) (Lane 3), U-87 MG treated with cobalt chloride (100 M for 16 hrs) (Lane 4), MDA-MB-231 (human breast adenocarcinoma cell line) (Lane 5), MDA-MB-231 treated with cobalt chloride (100 M for 16 hrs) (Lane 6), A549 (human lung carcinoma cell line) (Lane 7) and A549 treated with cobalt chloride (100 M for 16 hrs) (Lane 8).

The blots were probed with Anti-LOXL2 Recombinant Rabbit Multiclonal Antibody (ab277807, 2.5 μg/mL) and detected by chemiluminescence using a Goat anti-Rabbit IgG (H+L)-HRP secondary antibody, (0.4 μg/mL, 1/4000 dilution). A 87 kDa band corresponding to LOXL2 was observed across the cell lines tested.

4-12% Bis-Tris gel.

The membrane was probed with the relevant primary and secondary antibody following blocking with 5% skimmed milk.

Chemiluminescent detection was ECL.

All lanes:

Western blot - Anti-LOXL2 antibody [7HCLC] (ab277807)

Predicted band size: 87 kDa

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• WB

Supplier Data

Western blot - Anti-LOXL2 antibody [7HCLC] (AB277807)

Knockdown of LOXL2 was achieved by transfecting U87MG cells with LOXL2 specific validated siRNA.

Western blot analysis was performed using modified whole cell extract (1% SDS) from the LOXL2 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and nontransfected cells (lane 1).

The blots were probed with ab277807 at 1-3 μg/mL and a Goat anti-Rabbit IgG (H+L)-HRP secondary antibody (0.4 μg/mL, 1/2500 dilution).

Loss of signal upon siRNA mediated knock down confirms that antibody is specific to LOXL2.

All lanes:

Western blot - Anti-LOXL2 antibody [7HCLC] (ab277807)

Predicted band size: 87 kDa

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