Anti-LOXL4 antibody [EPR28300-90]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-LOXL4 antibody [EPR28300-90] (AB313797)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell, Left) / A375 (human malignant melanoma epithelial cell, Right) cells labelling LOXL4 with ab313797 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Low expression : HeLa (PMID : 30728460).

• IP

Supplier Data

Immunoprecipitation - Anti-LOXL4 antibody [EPR28300-90] (AB313797)

LOXL4 was immunoprecipitated from 0.35 mg A375 (human malignant melanoma epithelial cell) whole cell lysate with ab313797 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313797 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : A375 (human malignant melanoma epithelial cell) whole cell lysate Lane 2 : A375 (human malignant melanoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313797 in A375 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-LOXL4 antibody [EPR28300-90] (ab313797) at 1/30 dilution

All lanes:

A375 (human malignant melanoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

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• WB

Supplier Data

Western blot - Anti-LOXL4 antibody [EPR28300-90] (AB313797)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : LoVo, HeLa(PMID : 30728460). The molecular weight observed is consistent with what has been described in the literature (PMID : 30728460). This blot of Lane 4 was developed using a high sensitivity ECL substrate. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : Lane1-3 : 15 seconds, Lane 4 : 180 seconds.

All lanes:

Western blot - Anti-LOXL4 antibody [EPR28300-90] (ab313797) at 1/1000 dilution

Lane 1:

A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

Huh7 (human hepatocellar carcinoma 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

Observed band size: 90 kDa

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• WB

Supplier Data

Western blot - Anti-LOXL4 antibody [EPR28300-90] (AB313797)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : NIH/3T3(PMID : 11292829).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-LOXL4 antibody [EPR28300-90] (ab313797) at 1/1000 dilution

Lane 1:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 30 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 30 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

Observed band size: 90 kDa

true

Exposure time: 125s