重组Anti-Cyclin B1抗体[EPR17060]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Cyclin B1 antibody [EPR17060] (AB181593)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Cyclin B1 with ab181593 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B1 antibody [EPR17060] (AB181593)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Cyclin B1 with ab181593 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasm and weak nuclear staining on HeLa 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 1/1000 dilution (red).
The negative controls are as follows :
1. ab181593 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [EPR17060] (AB181593)

Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling Cyclin B1 using ab181593 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Negative staining on Human brain tissue. Counter stained with Hematoxylin.

Negative 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [EPR17060] (AB181593)

Immunohistochemical analysis of paraffin-embedded Human lung squamous cell carcinomal tissue labeling Cyclin B1 using ab181593 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Cytoplasm staining on cancer cells of Human lung squamous cell carcinoma is observed. Counter stained with Hematoxylin.

Negative 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [EPR17060] (AB181593)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Cyclin B1 using ab181593 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Cytoplasm staining on the germinal center of Human tonsil is observed. Counter stained with Hematoxylin.

Negative 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

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Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B1 antibody [EPR17060] (AB181593)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 (Mouse myoblast cell line) cells labeling Cyclin B1 with ab181593 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasm and nuclear staining on C2C12 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 1/1000 dilution (red).
The negative controls are as follows :
1. ab181593 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [EPR17060] (AB181593)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Cyclin B1 using ab181593 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Cytoplasm staining on epithelial cells of mouse colon is observed. Counter stained with Hematoxylin.

Negative 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [EPR17060] (AB181593)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Cyclin B1 using ab181593 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Negative staining on Mouse liver tissue. Counter stained with Hematoxylin.

Negative 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.

• WB

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Western blot - Anti-Cyclin B1 antibody [EPR17060] (AB181593)

5% NFDM/TBST : Blocking and diluting buffer.

All lanes:

Western blot - Anti-Cyclin B1 antibody [EPR17060] (ab181593) at 1/2000 dilution

Lane 1:

C2C12 (Mouse myoblast cell line) whole cell lysate at 10 µg

Lane 2:

NIH 3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/1000 dilution

Predicted band size: 48 kDa

Observed band size: 55 kDa

false

Exposure time: 30s

• WB

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Western blot - Anti-Cyclin B1 antibody [EPR17060] (AB181593)

5% NFDM/TBST : Blocking and diluting buffer.

All lanes:

Western blot - Anti-Cyclin B1 antibody [EPR17060] (ab181593) at 1/10000 dilution

Lane 1:

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

Lane 2:

Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/1000 dilution

Predicted band size: 48 kDa

Observed band size: 55 kDa

false

Exposure time: 30s

• WB

Lab

Western blot - Anti-Cyclin B1 antibody [EPR17060] (AB181593)

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : CCNB1 (KO) knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : Hek293 whole cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab181593 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab181593 was shown to recognize CCNB1 when CCNB1 knockout samples were used, along with additional cross-reactive bands. Wild-type and CCNB1 (KO) knockout samples were subjected to SDS-PAGE. ab181593 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 2000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

Predicted band size: 48 kDa

false