Anti-LOX antibody [EPR4025]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-LOX antibody [EPR4025] (AB174316)

Immunofluorescence staining of Jurkat cells with purified ab174316 at a working dilution of 1 in 300 counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti rabbit used at a dilution of 1 in 200. The cells were fixed in 4% PFA.

• ICC/IF

AbReview82508****

Immunocytochemistry/ Immunofluorescence - Anti-LOX antibody [EPR4025] (AB174316)

Immunocytochemistry/ Immunofluorescence analysis of formaldehyde-fixed Triton X-100 permeabilized human chondrocytes staining with ab174316 at 1/100 dilution in PBS with 0.1% triton x100. Secondary antibody was Alexa Fluor® 488 Goat Anti-Rabbit at 1/2000 dilution. Samples were incubated with the primary antibody for 30 minutes at 37°C. Blocking was with 0.1% BSA for 30 minutes at 37°C (ref : PMC7204390)

This image is courtesy of an anonymous Abreview

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LOX antibody [EPR4025] (AB174316)

Immunohistochemical staining of paraffin embedded human kidney with purified ab174316 at a working dilution of 1 in 900. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer pH 9.0.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-LOX antibody [EPR4025] (AB174316)

Flow Cytometry (Intracellular) analysis of ab174316. HeLa (Human cervix adenocarcinoma epithelial cell) cells were fixed in 4% paraformaldehyde and permeabilized with 90% methanol. ab174316 was used at 1 : 100 dilution (1ug) (Red). Secondary antibody Goat anti rabbit IgG (Alexa Fluor® 647, ab150083) at 1 : 5000 dilution. Isotype control Rabbit monoclonal IgG (ab172730) (Black). Unlabelled control, cells without incubation with primary antibody and secondary antibody (Blue).

• IP

Supplier Data

Immunoprecipitation - Anti-LOX antibody [EPR4025] (AB174316)

Western blot analysis on immunoprecipitation pellet from (Lane 1) WI-38 (Human fetal lung fibroblast cell line) cells lysate or (Lane 2) 1X PBS (negative control) using unpurified ab174316 at a 1/10 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

All lanes:

Immunoprecipitation - Anti-LOX antibody [EPR4025] (ab174316)

Predicted band size: 47 kDa

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• WB

Lab

Western blot - Anti-LOX antibody [EPR4025] (AB174316)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-LOX antibody [EPR4025] (ab174316) at 1/3700 dilution

Lane 1:

WI-38 cell lysate at 20 µg

Lane 2:

Jurkat cell lysate at 20 µg

Lane 3:

Mouse brain at 20 µg

Lane 4:

Rat brain at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 47 kDa

Observed band size: 50 kDa

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• WB

Lab

Western blot - Anti-LOX antibody [EPR4025] (AB174316)

False colour image of Western blot : Anti-LOX antibody [EPR4025] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab174316 was shown to bind specifically to LOX. A band was observed at 52 kDa in wild-type HeLa cell lysates with no signal observed at this size in Lox knockout cell line ab261801 (knockout cell lysate ab256981). To generate this image, wild-type and Lox knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-LOX antibody [EPR4025] (ab174316) at 1/1000 dilution

Lane 1:

Wild-type HeLa Vehicle Control DFO (0 mM, 24 h) cell lysate at 20 µg

Lane 2:

Wild-type HeLa Treated DFO (0.5 mM, 24 h) cell lysate at 20 µg

Lane 2:

Western blot - Human LOX knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-lox-knockout-hela-cell-line-ab261801'>ab261801</a>)

Lane 3:

LOX knockout HeLa Vehicle Control DFO (0 mM, 24 h) cell lysate at 20 µg

Lane 4:

LOX knockout HeLa Treated DFO (0.5 mM, 24 h) cell lysate at 20 µg

Predicted band size: 47 kDa

Observed band size: 52 kDa

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• WB

Lab

Western blot - Anti-LOX antibody [EPR4025] (AB174316)

Primary antibody dilution :

ab174316 Anti-LOX antibody 1 : 5000 dilution (0.1mg/ml)

ab179483 Anti-HIF-1 alpha antibody 1 : 500 dilution (0.3mg/ml)

Blocking buffer / Dilution buffer and concentration :

5% NFDM/TBST

LOX and HIF-1 alpha could be induced under hypoxia condition (PMID : 23545606, PMID : 30226558).

All lanes:

Western blot - Anti-LOX antibody [EPR4025] (ab174316)

Lane 1:

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

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with 0.5 mM DFO for 24 hours whole cell lysates 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 1/10000 dilution

Predicted band size: 47 kDa

Observed band size: 50 kDa

false

Exposure time: 7s