Anti-Cyclin A2 antibody [E399] - BSA and Azide free

• Flow Cyt

Lab

Flow Cytometry - Anti-Cyclin A2 antibody [E399] - BSA and Azide free (AB247261)

This data was developed using ab247261, the same antibody clone in a different buffer formulation.
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Cyclin A2 with Purified ab247261 at 1 : 30 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor™ 488, ab150077) secondary antibody was used at 1 : 2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cyclin A2 antibody [E399] - BSA and Azide free (AB247261)

This data was developed using ab32498, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cyclin A2 with Purified ab32498 at 1/50 dilution (5.66 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Cyclin A2 antibody [E399] - BSA and Azide free (AB247261)

This data was developed using ab32498, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of cyclin A expression in HeLa cells using 1/250 ab32498.

• IP

Lab

Immunoprecipitation - Anti-Cyclin A2 antibody [E399] - BSA and Azide free (AB247261)

This data was developed using ab32498, the same antibody clone in a different buffer formulation.
Purified ab32498 at 1/20 dilution (1μg) immunoprecipitating Cyclin A2 in K-562 whole cell lysate.
Lane 1 (input) : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10μg
Lane 2 (+) : ab32498 + K-562 whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab32498 in K-562 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 50 kDa

All lanes:

Immunoprecipitation - Anti-Cyclin A2 antibody [E399] - BSA and Azide free (ab247261)

Predicted band size: 49 kDa

false

• WB

Supplier Data

Western blot - Anti-Cyclin A2 antibody [E399] - BSA and Azide free (AB247261)

The smaller band is due to a tissue-specific splice variant(PMID 22745723). The band of 35kDa maybe the cleaved form(PMID : 12176996).

All lanes:

Western blot - Anti-Cyclin A2 antibody [E399] (<a href='/en-us/products/primary-antibodies/cyclin-a2-antibody-e399-ab32498'>ab32498</a>) at 1/2000 dilution

Lane 1:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 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: 50 kDa

false

• WB

Supplier Data

Western blot - Anti-Cyclin A2 antibody [E399] - BSA and Azide free (AB247261)

Cyclin A2 is down regulated at the G2/M phase.

All lanes:

Western blot - Anti-Cyclin A2 antibody [E399] (<a href='/en-us/products/primary-antibodies/cyclin-a2-antibody-e399-ab32498'>ab32498</a>) at 1/1000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate; Untreated, asynchronous cells (<a href='/en-us/products/unavailable/hela-cell-cycle-lysates-thymidine-treated-nocodazole-treated-untreated-asynchronous-control-ab136811'>ab136811</a>) at 20 µg

Lane 2:

HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate; G1/S arrested cells (thymidine treatment) (<a href='/en-us/products/unavailable/hela-cell-cycle-lysates-thymidine-treated-nocodazole-treated-untreated-asynchronous-control-ab136811'>ab136811</a>) at 20 µg

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate; G2/M arrested cells (sequential thymidine and nocodazole treatments) (<a href='/en-us/products/unavailable/hela-cell-cycle-lysates-thymidine-treated-nocodazole-treated-untreated-asynchronous-control-ab136811'>ab136811</a>) 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: 50 kDa

false