MA1-156

antibody from Invitrogen Antibodies

Targeting: CCNB2 HsT17299

Provider product page for MA1-156

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [0]
Comments [0]
Validations
Western blot [3]
Immunocytochemistry [5]
Immunohistochemistry [3]
Other assay [1]

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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.
Antibody clone number: X29.2
Concentration: 1 mg/mL

CCNB2 protein structure - MA1-156 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Cyclin B2 was performed by loading 10 µg of HeLa and HEK293T cell lysates from cells left untreated or treated with 2mM hydroxyurea, or 100 nM camptothecin for 16 hours per lane onto a 4-12% Bis-Tris polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA in TBST for at least 1 hour. The membrane was probed with a Cyclin B2 monoclonal antibody (Product # MA1-156) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBST, and probed with an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Cyclin B2 was performed by loading 10 µg of COS7 cell lysates from cells left untreated or treated with 2mM hydroxyurea or 100 nM camptothecin for 16 hours per lane onto a 4-12% Bis-Tris polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA in TBST for at least 1 hour. The membrane was probed with a Cyclin B2 monoclonal antibody (Product # MA1-156) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBST, and probed with an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extract (30 µg lysate) of HeLa (Lane 1), HeLa treated with Doxorubicin (0.5uM, 24 hrs) (Lane 2), MCF7 (Lane 3), MCF7 treated with Nocodazole (50ng/ml, 24 hrs) (Lane 4), COS-7 (Lane 5), A431 (Lane 6) and Jeko1(Lane 7). The blot was probed with Anti-Cyclin B2 Monoclonal Antibody (X29.2) (Product # MA1-156, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A 52 kDa band corresponding to Cyclin B2 was observed in all cell lines tested and was enhanced upon treatments.

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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cyclin B2 showing staining in the cytoplasm of paraffin-embedded human cervical carcinoma (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Cyclin B2 monoclonal antibody (Product # MA1-156) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cyclin B2 showing staining in the cytoplasm of paraffin-embedded human colon tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Cyclin B2 monoclonal antibody (Product # MA1-156) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cyclin B2 showing staining in the nucleus and cytoplasm of paraffin-embedded mouse colon tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Cyclin B2 monoclonal antibody (Product # MA1-156) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.