Anti-Cyclin A2 antibody [Y193]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Cyclin A2 antibody [Y193] (AB32386)

Immunofluorescence staining of MCF7 cells with purified ab32386 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4 % PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab32386 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin A2 antibody [Y193] (AB32386)

Immunohistochemical staining of paraffin embedded human colon carcinoma with purified ab32386 at a working dilution of 1/500. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IP

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Immunoprecipitation - Anti-Cyclin A2 antibody [Y193] (AB32386)

ab32386 (purified) at 1/40 immunoprecipitating Cyclin A2 in 10 μg HeLa cell lysate (Lanes 1 and 2, observed at 55 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, HRP Veriblot for IP Detection Reagent (ab131366) was used for detection (1/10 000). Blocking buffer and concentration : 5% NFDM/TBST Dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-Cyclin A2 antibody [Y193] (ab32386)

Predicted band size: 49 kDa

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• WB

Lab

Western blot - Anti-Cyclin A2 antibody [Y193] (AB32386)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Cyclin A2 antibody [Y193] (ab32386) at 1/10000 dilution

Lane 1:

HepG2 cell lysate at 20 µg

Lane 2:

HEK293 cell 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 1/20000 dilution

Predicted band size: 49 kDa

Observed band size: 55 kDa

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• WB

Lab

Western blot - Anti-Cyclin A2 antibody [Y193] (AB32386)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Cyclin A2 antibody [Y193] (ab32386) at 1/10000 dilution

All lanes:

NIH/3T3 cell 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 1/20000 dilution

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• WB

Unknown

Western blot - Anti-Cyclin A2 antibody [Y193] (AB32386)

All lanes:

Western blot - Anti-Cyclin A2 antibody [Y193] (ab32386) at 1/1000 dilution

All lanes:

HeLa cell lysate

Predicted band size: 49 kDa

Observed band size: 55 kDa

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