Anti-Superoxide Dismutase 1 antibody [EP1727Y]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (AB51254)

Immunofluorescent staining of HeLa cells using ab51254 (1 : 100).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (AB51254)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Superoxide Dismutase 1 with ab51254 at 1/20 dilution (1μg) (Red). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Cells were fixed with4% paraformaldehyde. Rabbit monoclonal IgG (ab172730) was used as isotype control (Black). Unlabelled control : Cells without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (AB51254)

ab51254 (1 : 100) staining human Superoxide Dismutase 1 in human breast carcinoma tissue by immunohistochemistry using paraffin embedded tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (AB51254)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-12 cells labelling Superoxide Dismutase 1 with ab51254 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in PC-12 cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 2.5 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 2 ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (AB51254)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling Superoxide Dismutase 1 with purified ab51254 at 1/1000 dilution (0.15 µg/ml).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : secondary antibody only control. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (AB51254)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling Superoxide Dismutase 1 with purified ab51254 at 1/1000 dilution (0.15 µg/ml).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : secondary antibody only control. Hematoxylin was used as a counterstain.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (AB51254)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling Superoxide Dismutase 1 with ab51254 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic and weak nuclear staining in NIH/3T3 cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 2.5 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 2 ug/ml dilution.

• WB

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Western blot - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (AB51254)

All lanes:

Western blot - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (ab51254) at 1/50000 dilution

All lanes:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP labeled at 1/2000 dilution

Predicted band size: 15 kDa

Observed band size: 18 kDa

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• WB

Collaborator

Western blot - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (AB51254)

ab51254 was shown to react with SOD1 in wild-type HeLa cells in Western blot with loss of signal observed in a SOD1 knockout cell line. Wild-type HeLa and SOD1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab51254 overnight at 4 °C at a 1/15000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2 μg/ml before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (ab51254)

Predicted band size: 15 kDa

false

• WB

Unknown

Western blot - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (AB51254)

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

All lanes:

Western blot - Anti-Superoxide Dismutase 1 antibody [EP1727Y] (ab51254) at 0.015 µg/mL

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate 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 0.05 µg/mL

Predicted band size: 15 kDa

Observed band size: 18 kDa

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