Anti-CDKN1B/p27 KIP 1 antibody (bs-0742R)

Anti-CDKN1B/p27 KIP 1 antibody

DATASHEET

Host:Rabbit

Target Protein:CDKN1B/p27 KIP 1

IR:Immunogen Range:101-198/198

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:1027

Swiss Prot:P46527

Source:KLH conjugated synthetic peptide derived from human P27 kip1:101-198/198 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:Cell cycle progression is regulated by cyclins and their cognate Cdks. p27 KIP 1 is a cell cycle regulatory mitotic inhibitor of cdk activity. p27 KIP 1 is a candidate tumor suppressor gene, and has been proposed to function as a possible mediator of TGF beta induced G1 arrest. p27 KIP 1 is up regulated in response to antimitogenic stimuli. The increased protein expression of p27 results in cellular arrest by binding to cyclin/Cdk complexes such as cyclin D1/Cdk4. p27 Kip1 is regulated by phosphorylation on serine 10 (S10) and threonine 187 (T187). Phosphorylation by CDK2 on T187 results in ubiquitylation and degradation of p27 Kip 1; while phosphorylation by hKIS on S10 signals the nuclear export to the cytoplasm.

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:5000-10000)
IP(1:20-100)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(1μg/Test )
ICC(1:100)
IF(1:200-800)

Cross Reactive Species:Human
Mouse
Rat
Chicken
Dog
Pig
Sheep
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Sample: Hela Cell (Human) Lysate at 40 ug
Primary: Anti-CDKN1B/p27 KIP 1 (bs-0742R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 22 kD
Observed band size: 24 kD

CDKN1B (p27Kip1) was immunoprecipitated from mouse kidney tissue with bs-0742R at 1/150 dilution. Western blot was performed from the immunoprecipitate using protein A/G beads. HRP Conjugated Goat anti-Rabbit IgG (Heavy Chain specific) was used as secondary antibody at 1:5000 dilution.
Lane 1: mouse kidney tissue lysate 10 μg (Input).
Lane 2: bs-0742R IP in mouse kidney tissue lysate.
Lane 3: native rabbit IgG IP in mouse kidney tissue lysate (negative control).
Secondary
All lanes Goat anti-Rabbit IgG (Heavy Chain specific), HRP Conjugated, 1:5000

Sample:
293T(Human) Cell Lysate at 30 ug
Primary: Anti- CDKN1B'p27 KIP 1 (bs-0742R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 22 kD
Observed band size: 22 kD

Sample:
293T(Human) Cell Lysate at 30 ug
U2os(Human) Cell Lysate at 30 ug
Primary: Anti- CDKN1B’p27 KIP 1 (bs-0742R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 22 kD
Observed band size: 22 kD

Sample:
Brain (Mouse) Lysate at 40 ug
Primary: Anti- CDKN1B'p27 KIP 1 (bs-0742R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 22 kD
Observed band size: 22 kD

Tissue/cell: human ovary carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37∩ for 20 min;
Incubation: Anti-CDKN1B/P27kip1 Polyclonal Antibody, Unconjugated(bs-0742R) 1:200, overnight at 4∑C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Tissue/cell: MCF7; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (CDKN1B ) Polyclonal Antibody, Unconjugated (bs-0742R) 1:200, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Paraformaldehyde-fixed, paraffin embedded (Human glioma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation: Anti-CDKN1B p27 KIP 1 Antibody, conjugated (bs-0742R-PE-Cy7) 1:400, 1.5 hours at 37°C; DAPI (5ug/ml, blue, C-0033) was used to stain the cell nuclei.