Anti-Cyclin B2/CCNB2 antibody [R17985] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B2/CCNB2 antibody [R17985] - BSA and Azide free (AB250841)

This data was developed using ab185622, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SW480 (Human colon adenocarcinoma cell line) cells labeling Cyclin B2/CCNB2 with ab185622 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic staining on SW480 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
-ve control 1 : ab185622 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B2/CCNB2 antibody [R17985] - BSA and Azide free (AB250841)

This data was developed using ab185622, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling Cyclin B2/CCNB2 with ab185622 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic staining on tumor cells of Human colon epithelium is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B2/CCNB2 antibody [R17985] - BSA and Azide free (AB250841)

This data was developed using ab185622, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Cyclin B2/CCNB2 with ab185622 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic staining on normal Human colon epithelium is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-Cyclin B2/CCNB2 antibody [R17985] - BSA and Azide free (AB250841)

This data was developed using ab185622, the same antibody clone in a different buffer formulation.

Cyclin B2/CCNB2 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab185622 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab185622 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : HeLa whole cell lysate 10 μg (Input).
Lane 2 : ab185622 IP in HeLa whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab185622 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds

All lanes:

Immunoprecipitation - Anti-Cyclin B2/CCNB2 antibody [R17985] (<a href='/en-us/products/primary-antibodies/cyclin-b2-ccnb2-antibody-r17985-ab185622'>ab185622</a>)

Predicted band size: 45 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B2/CCNB2 antibody [R17985] - BSA and Azide free (AB250841)

This data was developed using ab185622, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling Cyclin B2/CCNB2 with ab185622 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic staining on spermatocytes of rat testis is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B2/CCNB2 antibody [R17985] - BSA and Azide free (AB250841)

This data was developed using ab185622, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 (Rat adrenal gland pheochromocytoma) cells labeling Cyclin B2/CCNB2 with ab185622 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic staining on PC-12 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
-ve control 1 : ab185622 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• WB

Supplier Data

Western blot - Anti-Cyclin B2/CCNB2 antibody [R17985] - BSA and Azide free (AB250841)

This data was developed using ab185622, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Cyclin B2/CCNB2 antibody [R17985] (<a href='/en-us/products/primary-antibodies/cyclin-b2-ccnb2-antibody-r17985-ab185622'>ab185622</a>) at 1/1000 dilution

Lane 1:

SW480 (Human colon adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Cyclin B2/CCNB2 antibody [R17985] - BSA and Azide free (AB250841)

This data was developed using ab185622, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Cyclin B2/CCNB2 antibody [R17985] (<a href='/en-us/products/primary-antibodies/cyclin-b2-ccnb2-antibody-r17985-ab185622'>ab185622</a>) at 1/5000 dilution

Lane 1:

A549 (Human lung carcinoma) whole cell lysate at 20 µg

Lane 2:

Mouse testis lysate at 20 µg

Lane 3:

Rat testis 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 1/1000 dilution

Predicted band size: 24 kDa,45 kDa

Observed band size: 24 kDa,45 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Cyclin B2/CCNB2 antibody [R17985] - BSA and Azide free (AB250841)

This data was developed using ab185622, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Cyclin B2/CCNB2 antibody [R17985] (<a href='/en-us/products/primary-antibodies/cyclin-b2-ccnb2-antibody-r17985-ab185622'>ab185622</a>) at 1/1000 dilution

Lane 1:

C6 (Rat glial tumor cells) whole cell lysate at 20 µg

Lane 2:

PC-12 (Rat adrenal gland pheochromocytoma) whole 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 1/1000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

Exposure time: 30s