重组Anti-LOX抗体[EPR4025] - BSA and Azide free (ab271926)

All lanes : Anti-LOX antibody [EPR4025] (ab174316) at 1/1000 dilution

Lane 1 : Wild-type HeLa Vehicle Control DFO (0 mM, 24 h) cell lysate
Lane 2 : Wild-type HeLa Treated DFO (0.5 mM, 24 h) cell lysate
Lane 3 : LOX knockout HeLa Vehicle Control DFO (0 mM, 24 h) cell lysate
Lane 4 : LOX knockout HeLa Treated DFO (0.5 mM, 24 h) cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 47 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?

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).

Overlay histogram showing Jurkat cells fixed in 2% PFA and stained with purified ab174316 at a dilution of 1 in 90 (pink line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 150. Rabbit monoclonal IgG was used as an isotype control. 

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

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

Immunofluorescence staining of Jurkat cells with unpurified ab174316 at a working dilution of 1 in 100, 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).

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).

Immunohistochemical staining of paraffin embedded human kidney with unpurified ab174316 at a working dilution of 1 in 300. 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).

Immunohistochemical analysis of paraffin-embedded human muscle tissue labeling LOX with unpurified ab174316 at a 1/50 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

Immunofluorescence analysis of HeLa cells labeling LOX with unpurified ab174316 at a 1/50 dilution.

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

Intracellular flow cytometric analysis of permeabilized Jurkat cells using unpurified ab174316 at a 1/10 dilution  (red) or a rabbit IgG (negative) (green). 

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

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