MA1-156

antibody from Invitrogen Antibodies

Targeting: CCNB2 HsT17299

Western blot

Immunoprecipitation

Immunohistochemistry

Antibody data

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Validation data

Product number: MA1-156 - Provider product page
Provider: Invitrogen Antibodies
Product name: Cyclin B2 Monoclonal Antibody (X29.2)
Antibody type: Monoclonal
Antigen: Other
Description: MA1-156 detects Cyclin B2 in mammalian samples. MA1-156 has been successfully used in Immunofluorescence, Immunoprecipitation, IHC (P) and Western Blot procedures. Immunoprecipitation and Western Blot analysis with MA1-156 show the accumulation of a prominent band at ~51 kDa in camptothecin and hydroxyurea treated cells. MA1-156 also detects additional unknown band at ~80 kDa. In Immunofluorescence applications, MA1-156 shows cyclin B2 staining consistent with the Golgi region, whereas MA1-155 shows cyclin B1 co-localization with microtubules. MA1-156 reacts with Cyclin B2 from Xenopus laevis and mammalian sources. In Western blot applications, X29.2 also cross reacts with Cyclin B1.
Reactivity: Human, Mouse, Xenopus
Host: Mouse
Isotype: IgG
Antibody clone number: X29.2
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: -20°C

CCNB2 protein structure - MA1-156 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Cyclin B2 (green) showing staining in the in the cytoplasm of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cyclin B2 monoclonal antibody (Product # MA1-156) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Cyclin B2 (green) in asynchronous HeLa cells showing cytoplasmic and nuclear localization. Images shown are without (left panel) or with (right panel) nuclear Hoechst staining. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 15 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a Cyclin B2 monoclonal antibody (Product # MA1-156) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502). Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249) for 30 minutes. Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Cyclin B2 (green) showing staining in the in the cytoplasm of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cyclin B2 monoclonal antibody (Product # MA1-156) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Cyclin B2 (green) showing staining in the in the cytoplasm of SW480 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cyclin B2 monoclonal antibody (Product # MA1-156) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Cyclin B2 was performed using 70% confluent log phase HeLa cells treated with Camptothecin (100nM, 16 hrs). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cyclin B2 Monoclonal Antibody (Product # MA1-156) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing accumulation of Cyclin A2 in the nucleus upon treatment. Panel e represents the control cells showing cytoplasmic staining. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.