Anti-LOX antibody [EPR4025] - Low endotoxin, Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LOX antibody [EPR4025] - Low endotoxin, Azide free (AB219369)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab174316).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-LOX antibody [EPR4025] - Low endotoxin, Azide free (AB219369)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab174316).

• IP

Supplier Data

Immunoprecipitation - Anti-LOX antibody [EPR4025] - Low endotoxin, Azide free (AB219369)

Western blot analysis on immunoprecipitation pellet from (Lane 1) WI-38 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab174316).

All lanes:

Immunoprecipitation - Anti-LOX antibody [EPR4025] (<a href='/en-us/products/primary-antibodies/lox-antibody-epr4025-ab174316'>ab174316</a>)

Predicted band size: 47 kDa

false

• WB

Lab

Western blot - Anti-LOX antibody [EPR4025] - Low endotoxin, Azide free (AB219369)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab174316).

All lanes:

Western blot - Anti-LOX antibody [EPR4025] (<a href='/en-us/products/primary-antibodies/lox-antibody-epr4025-ab174316'>ab174316</a>) 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

false