Cyclin B1 Rabbit Polyclonal Antibody

• Catalog Number: orb10494
• Category: Antibodies
• Description: Cyclin B1 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Cyclin B1 (271-433/433aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 48 kDa
• Target: CCNB1
• UniProt ID: P14635
• RRID: AB_10748485
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: CCNB 1; CCNB; CCNB1; CCNB1_HUMAN; G2 mitotic speci
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Wang, Qi et al. Methamphetamine induces hepatotoxicity via inhibiting cell division, arresting cell cycle and activating apoptosis: In vivo and in vitro studies Food Chem Toxicol, 105, 61-72 (2017)

PubMed: 28341135

Blank control (blue line): A549 (blue). Primary Antibody (green line): Rabbit Anti-Cyclin B1 antibody (orb10494). 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. Acquisition of 20000 events was performed.

Cell: Hela, Concentration: 1:100, Host/Isotype: Rabbit/IgG, Flow cytometric analysis of primary antibody (Cat#: orb10494) 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.

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 (orb10494) 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 nuclei.

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Cyclin B1) Polyclonal Antibody, Unconjugated (orb10494) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Cyclin B1) Polyclonal Antibody, Unconjugated (orb10494) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Lymph node (Rat) Lysate at 40 ug, Lane 2: Testis (Rat) Lysate at 40 ug, Lane 3: Spleen (Rat) Lysate at 40 ug, Primary: Anti-Cyclin B1 (orb10494) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 55-60 kD, Observed band size: 60 kD.

Sample: U251 Cell (Human) Lysate at 30 ug, Primary: Anti-Cyclin B1 (orb10494) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 48 kD, Observed band size: 50 kD.

Sample: U937 Cell (Human) Lysate at 30 ug, Primary: Anti-Cyclin B1 (orb10494) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 48 kD, Observed band size: 50 kD.

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 (orb10494) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.