phospho-CDKN1B (Thr157) Polyclonal Antibody

$MCF-7 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with CDKN1B (Thr157) Polyclonal Antibody(bs-5228R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

MCF-7 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with CDKN1B (Thr157) Polyclonal Antibody(bs-5228R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

$Paraformaldehyde-fixed, paraffin embedded Mouse muscle; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with CDKN1B (Thr157) Polyclonal Antibody, Unconjugated (bs-5228R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse muscle; 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\u00b0C for 30min; Antibody incubation with CDKN1B (Thr157) Polyclonal Antibody, Unconjugated (bs-5228R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Rat muscle; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with CDKN1B (Thr157) Polyclonal Antibody, Unconjugated (bs-5228R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat muscle; 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\u00b0C for 30min; Antibody incubation with CDKN1B (Thr157) Polyclonal Antibody, Unconjugated (bs-5228R) at 1:200 overnight at 4\u00b0C, DAB staining.

Lane 1: Mouse Spleen lysates; Lane 2: Mouse Thymus lysates; Lane 3: Rat Thymus lysates probed with CDKN1B Polyclonal Antibody, Unconjugated (bs-5228R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Applications

WB
IHC-P
IHC-F
IF
Flow-Cyt

Reactivity

Human
Mouse
Rat

Predicted Reactivity

Dog
Pig
Rabbit
Guinea Pig

Overview
Catalog #	bs-5228R
Product Name	phospho-CDKN1B (Thr157) Polyclonal Antibody
Applications	WB, IHC-P, IHC-F, IF, Flow-Cyt
Reactivity	Human, Mouse, Rat
Predicted Reactivity	Dog, Pig, Rabbit, Guinea Pig
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic phosphopeptide derived from human CDKN1B around the phosphorylation site of Thr157 [PA(p-T)DD]
Modification Site	Thr157
Clonality	Polyclonal
Clone #	#REF!
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4_. Store at -20_ for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	1027
Subcellular location	Cytoplasm, Nucleus
Synonyms	p27Kip1Thr157; p-p27Kip1Thr157; p-p27Kip1T157; CDN1B_HUMAN; AA408329; AI843786; Cdki1b; CDKN 1B; CDKN 4; CDKN1B; CDKN1B; CDKN4; CDKN4; Cyclin Dependent Kinase Inhibitor 1B; Cyclin Dependent Kinase Inhibitor 1B; Cyclin dependent kinase inhibitor p27; Cyclin dependent kinase inhibitor p27; Cyclin-dependent kinase inhibitor 1B p27, Kip1; Cyclin-dependent kinase inhibitor 1B; Cyclin-dependent kinase inhibitor p27; Cyclin-dependent kinase inhibitor p27 Kip1; KIP 1; KIP1; MEN1B; MEN4; OTTHUMP00000195098; OTTHUMP00000195099; p27; p27 Kip1; P27-like cyclin-dependent kinase inhibitor; P27KIP1.
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.
Application Dilution
WB	1:300-5000
IHC-P	ELISA1:500-1000
IHC-F	FCM1:20-100
IF	IHC-P1:200-400
Flow-Cyt