Lanes 1-5 : Anti-ACOX1/AOX antibody [EPR19038] (ab184032) at 1/1000 dilution
Lanes 6-7 : Anti-ACOX1/AOX antibody [EPR19038] (ab184032) at 1/5000 dilution

Lane 1 : Mouse liver lysate
Lane 2 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 3 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 5 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate
Lane 6 : Mouse heart lysate
Lane 7 : Mouse kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 75 kDa
Observed band size: 50,72 kDa why is the actual band size different from the predicted?

Blocking/dilution buffer: 5% NFDM/TBST.

Exposure time Lane 1-5: 1 second; Lane 6: 30 seconds; Lane 7: 2 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID:) 8798738 and 17255948.

All lanes : Anti-ACOX1/AOX antibody [EPR19038] (ab184032) at 1/1000 dilution

Lane 1 : Rat brain lysate
Lane 2 : Rat heart lysate
Lane 3 : Rat liver lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 75 kDa
Observed band size: 50,72 kDa why is the actual band size different from the predicted?

Blocking/dilution buffer: 5% NFDM/TBST.

Exposure time Lane 1/2: 8 seconds;Lane 3: 1 second.

The expression profile observed is consistent with what has been described in the literature (PMID:) 8798738 and 17255948.

All lanes : Anti-ACOX1/AOX antibody [EPR19038] (ab184032) at 1/1000 dilution

Lane 1 : Human fetal liver lysate
Lane 2 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 75 kDa
Observed band size: 50,72 kDa why is the actual band size different from the predicted?


Exposure time: 3 minutes

Blocking/dilution buffer: 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID:) 8798738 and 17255948.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cell line) cells labeling ACOX1/AOX with ab184032 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on C6 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [EPR19038] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows:
-ve control 1: ab184032 at 1/250 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291  at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling ACOX1/AOX with ab184032 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [EPR19038] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows:
-ve control 1: ab184032 at 1/250 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast) cell line labeling ACOX1/AOX with ab184032 at 1/100 dilution (red) compared with a Rabbit IgG, monoclonal [EPR19038] - Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

ACOX1/AOX was immunoprecipitated from 1mg of NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with ab184032 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab184032 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: NIH/3T3 whole cell lysate 10µg (Input).
Lane 2: ab184032 IP in NIH/3T3 whole cell lysate.
Lane 3: Rabbit IgG, monoclonal [EPR19038] - Isotype Control (ab172730) instead of ab184032 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.