Phospho-MDM2 (Ser166) Polyclonal Antibody

$Paraformaldehyde-fixed, paraffin embedded human colon carcinoma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with phospho-MDM2 (Ser166) Polyclonal Antibody, Unconjugated (bs-3266R) at 1:500 overnight at 4\u00b0C, followed by a conjugated secondary for 20 minutes and DAB staining.$

Paraformaldehyde-fixed, paraffin embedded human colon carcinoma; 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 phospho-MDM2 (Ser166) Polyclonal Antibody, Unconjugated (bs-3266R) at 1:500 overnight at 4\u00b0C, followed by a conjugated secondary for 20 minutes and DAB staining.

$Molt4 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with MDM2 (Ser166) Antibody(bs-3266R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

Molt4 cells 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 MDM2 (Ser166) Antibody(bs-3266R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

Applications

WB
ELISA
FCM
IHC-P
IHC-F
IF(IHC-P)
IF(IHC-F)
IF(ICC)

Reactivity

Human

Predicted Reactivity

Mouse
Rat
Horse
Rabbit

Overview
Catalog #	bs-3266R
Product Name	Phospho-MDM2 (Ser166) Polyclonal Antibody
Applications	WB, ELISA, FCM, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC)
Reactivity	Human
Predicted Reactivity	Mouse, Rat, Horse, Rabbit
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic phosphopeptide derived from human MDM2 around the phosphorylation site of Ser166
Modification Site	Ser166
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	4193
Subcellular location	Cytoplasm, Nucleus
Synonyms	MDM2Ser166 ; MDM2 Ser166; Double minute 2 protein; Hdm 2; HDM2; MDM 2; Mdm2 transformed 3T3 cell double minute 2 p53 binding protein mouse binding protein 104kDa; MDM2BP; Mouse Double Minute 2; MTBP; Murine Double Minute Chromosome 2; Oncoprotein Mdm2; p53 Binding Protein Mdm2; Ubiquitin protein ligase E3 Mdm2.
Background	Inhibits TP53/p53- and TP73/p73-mediated cell cyclearrest and apoptosis by binding its transcriptional activation domain. Functions as a ubiquitin ligase E3, in the presence of E1 and E2, toward p53 and itself. Permits the nuclear export of p53 and targets it for proteasome-mediated proteolysis. Binds p53, p73, ARF(P14), ribosomal protein L5 and specifically to RNA. Can interact also with retinoblastoma protein(RB), E1A-associated protein EP300 and the E2F1 transcription factor. Forms a ternary complex with TP53/p53 and WWOX. Interacts with CDKN2AIP, MTBP, TRBG1 and USP7. Isoform Mdm2-F does not interact with TP53/p53. Interacts with PYHIN1. Interacts with, and ubiquitinates HIV-1 Tat. Belongs to the MDM2/MDM4 family.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
FCM	1:20-100
IHC-P	1:200-400
IHC-F	1:100-500
IF(IHC-P)	1:50-200
IF(IHC-F)	1:50-200
IF(ICC)	1:50-200