Anti-Catalase antibody [EPR20198] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Catalase antibody [EPR20198] - BSA and Azide free (AB223793)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed C2C12 (Mouse myoblast cell line) cells labeling Catalase with ab209211 at 1/60 dilution (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209211).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Catalase antibody [EPR20198] - BSA and Azide free (AB223793)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 (Mouse myoblast cell line) cells labeling Catalase with ab209211 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on C2C12 cell line. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209211).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Catalase antibody [EPR20198] - BSA and Azide free (AB223793)

ImmunoFluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Catalase with ab209211 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cell line. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209211).

• WB

Lab

Western blot - Anti-Catalase antibody [EPR20198] - BSA and Azide free (AB223793)

This WB data was generated using the same anti-Catalase antibody clone [EPR20198] in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (cat# ab209211).

Lanes 1 - 4 : Merged signal (red and green). Green - ab209211 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab209211 was shown to specifically react with CAT when CAT knockout samples were used. Wild-type and CAT knockout samples were subjected to SDS-PAGE. ab209211 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Catalase antibody [EPR20198] - Peroxisome Marker (<a href='/en-us/products/primary-antibodies/catalase-antibody-epr20198-peroxisome-marker-ab209211'>ab209211</a>) at 1/2000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

CAT knockout HAP1 whole cell lysate at 20 µg

Lane 3:

Hek293 whole cell lysate at 20 µg

Lane 4:

HepG2 whole cell lysate at 20 µg

Predicted band size: 60 kDa

false

• WB

Lab

Western blot - Anti-Catalase antibody [EPR20198] - BSA and Azide free (AB223793)

This data was developed using the same antibody clone in a different buffer formulation (ab209211).

Lanes 1-2 : Merged signal (red and green). Green - ab209211 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.

ab209211 Anti-Catalase antibody [EPR20198] was shown to specifically react with Catalase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265250 (knockout cell lysate ab256859) was used. Wild-type and Catalase knockout samples were subjected to SDS-PAGE. ab209211 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Catalase antibody [EPR20198] - Peroxisome Marker (<a href='/en-us/products/primary-antibodies/catalase-antibody-epr20198-peroxisome-marker-ab209211'>ab209211</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CAT knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CAT (Catalase) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cat-catalase-knockout-hela-cell-line-ab265250'>ab265250</a>)

Predicted band size: 60 kDa

Observed band size: 60 kDa

false