Anti-CCNB1 Polyclonal Antibody is a Rabbit antibody targeting CCNB1. Anti-CCNB1 Polyclonal Antibody can be used in FCM, ICC/IF, IF, IHC-Fr, IHC-P, WB.

IgG

Human,Mouse,Rat (predicted:Cow)

1. Paraformaldehyde-fixed, paraffin embedded (Rat esophageal); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Cyclin B1) Polyclonal Antibody, Unconjugated (TMAB-00316) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. Tissue/cell: human colon carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Cyclin B1 Polyclonal Antibody, Unconjugated (TMAB-00316) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Cell: Hela
Concentration:1:100
Host/Isotype: Rabbit/IgG
Flow cytometric analysis of primary antibody (Cat#: TMAB-00316) on Hela (green) compared with isotype control in the absence of primary antibody (blue) followed by Alexa Fluor 488-conjugated goat anti-rabbit Igg (H+L) secondary antibody.
4. Sample:
Lane 1: LympHnode (Rat) Lysate at 40 μg
Lane 2: Testis (Rat) Lysate at 40 μg
Lane 3: Spleen (Rat) Lysate at 40 μg
Primary: Anti-Cyclin B1 (TMAB-00316) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 55-60 kDa
Observed band size: 60 kDa
5. Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Cyclin B1) polyclonal Antibody, Unconjugated (TMAB-00316) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
6. Blank control (blue line): A549 (blue).
Primary Antibody (green line): Rabbit Anti-Cyclin B1 antibody (TMAB-00316).
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): f (ab/)2 fragment goat anti-rabbit IgG-FITC.
Dilution: 1 μg/test.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
7. Sample:
U251 Cell (Human) Lysate at 30 μg
Primary: Anti-Cyclin B1 (TMAB-00316) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 50 kDa
8. Sample:
U937 Cell (Human) Lysate at 30 μg
Primary: Anti-Cyclin B1 (TMAB-00316) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 50 kDa
9. Paraformaldehyde-fixed, paraffin embedded (Mouse small intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Cyclin B1) Polyclonal Antibody, Unconjugated (TMAB-00316) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

FCM=1 μg/Test; ICC/IF=1:100-500; IF=1:100-500; IHC-Fr=1:100-500; IHC-P=1:100-500; WB=1:500-2000

Polyclonal

KLH conjugated synthetic peptide: human Cyclin B1

Human

CCNB1

G2/mitotic-specific cyclin-B1

Cyclin B family,Cyclins,Cyclin B Family,Cyclin B Family

Essential for the control of the cell cycle at the G2/M (mitosis) transition.