Phospho-SMAD1 (Ser463/465)/ SMAD5 (Ser463/465)/ SMAD9 (Ser465/467) (D5B10) Rabbit mAb #13820

Filter:

WB
IP
IF
F
ChIP

Supporting Data

REACTIVITY	H M R
SENSITIVITY	Endogenous
MW (kDa)	60
Source/Isotype	Rabbit IgG

Application Key:

WB-Western Blotting
IP-Immunoprecipitation
IF-Immunofluorescence
F-Flow Cytometry
ChIP-Chromatin Immunoprecipitation

Species Cross-Reactivity Key:

H-Human
M-Mouse
R-Rat

Product Information

Product Usage Information

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 10⁶ cells) per IP. This antibody has been validated using SimpleChIP^® Enzymatic Chromatin IP Kits.

Application	Dilution
Western Blotting	1:1000
Immunoprecipitation	1:50
Immunofluorescence (Immunocytochemistry)	1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized)	1:200 - 1:800
Chromatin IP	1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #67959.

Protocol

Specificity / Sensitivity

Phospho-SMAD1 (Ser463/465)/ SMAD5 (Ser463/465)/ SMAD9 (Ser465/467) (D5B10) Rabbit mAb recognizes endogenous levels of SMAD1 and SMAD5 protein when phosphorylated at Ser463/465 and SMAD9 (SMAD8) protein when phosphorylated at Ser465/467.

Species Reactivity:

Human, Mouse, Rat

The antigen sequence used to produce this antibody shares 100% sequence homology with the species listed here, but reactivity has not been tested or confirmed to work by CST. Use of this product with these species is not covered under our Product Performance Guarantee.

Species predicted to react based on 100% sequence homology:

Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser463/465 of human SMAD1 and SMAD5 protein.

Background

Bone morphogenetic proteins (BMPs) constitute a large family of signaling molecules that regulate a wide range of critical processes including morphogenesis, cell-fate determination, proliferation, differentiation, and apoptosis (1,2). BMP receptors are members of the TGF-β superfamily of Ser/Thr kinase receptors. Ligand binding induces multimerization, autophosphorylation, and activation of these receptors (3-5). They subsequently phosphorylate SMAD1 at Ser463 and Ser465 in the carboxy-terminal motif SSXS, as well as SMAD5 and SMAD9 (SMAD8) at their corresponding sites. These phosphorylated SMADs dimerize with the coactivating SMAD4 and translocate to the nucleus, where they regulate the transcription of target genes (5). MAP kinases and CDKs 8 and 9 are also reported to phosphorylate residues in the linker region of SMAD1, including Ser206. Phosphorylation of SMAD1 at Ser206 recruits Smurf1 to the linker region and leads to the degradation of SMAD1 (6). Phosphorylation at this site also promotes SMAD1 transcriptional activity by recruiting YAP to the linker region (7).

Database Links

UniProt ID: Q99717 , Q15797 , O15198

Entrez-Gene Id: 4090 , 4086 , 4093

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受，否书均被拒书，并且无效。

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For Research Use Only. Not For Use In Diagnostic Procedures.

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