All lanes : Anti-Superoxide Dismutase 1 antibody [EPR23549-163] (ab252426) at 1/1000 dilution

Lane 1 : Human brain tissue lysate
Lane 2 : Human placenta tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 15 kDa
Observed band size: 15 kDa

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

Exposure time: 5.5 seconds

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Superoxide dismutase 1 with ab252426 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human kidney (PMID: 29861522). The section was incubated with ab252426 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MCF7 cells labelling Superoxide dismutase 1 with ab252426 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplamic and nuclear staining in MCF7 cell line Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (Human breast adenocarcinoma epithelial cell) cells labelling Superoxide dismutase 1 with ab252426 at 1/500 dilution (0.1ug) (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, ab150077) at 1/2000 dilution was used as the secondary antibody.

Superoxide dismutase 1 was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab252426 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab252426 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 ug

Lane 2: ab252426 IP in MCF7 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252426 in MCF7 whole cell lysate

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

Exposure time: 3 seconds

All lanes : Anti-Superoxide Dismutase 1 antibody [EPR23549-163] (ab252426) at 1/1000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : A431 (human epidermoid carcinoma epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 15 kDa
Observed band size: 15 kDa

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

The expression profile and molecular weight observed is consistent with what has been described in the literature (PMID: 14526083).

Exposure time: 5.5 seconds