SMAD3 (Phospho-S423) Rabbit Polyclonal Antibody

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SMAD3 (Phospho-S423) Rabbit Polyclonal Antibody

Key features and details

Rabbit polyclonal antibody to SMAD3 (Phospho-S423)

Target: SMAD3 (Phospho-S423)
Source/Host: Rabbit
Reactivity: Human, Mouse, Rat, Bovine, Chicken, Monkey, Pig, Sheep, Zebrafish
Clonality: Polyclonal
Applications: WB, IF/ICC
Conjugation: Unconjugated
Storage: at-20°C

Brand:

CAT.NO. : APA09533

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Size:

50μL 100μL

Trail, Bulk size or Custom requests Please contact us

Product Details

Background

Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD3/SMAD4 complex, activates transcription. Also can form a SMAD3/SMAD4/JUN/FOS complex at the AP-1/SMAD site to regulate TGF-beta-mediated transcription. Has an inhibitory effect on wound healing probably by modulating both growth and migration of primary keratinocytes and by altering the TGF-mediated chemotaxis of monocytes. This effect on wound healing appears to be hormone-sensitive. Regulator of chondrogenesis and osteogenesis and inhibits early healing of bone fractures.

Application

To ensure optimal assay performance, AREX recommends conducting reagent titration tailored to each testing system for optimal detection results.

WB	1:500 - 1:1000
IF/ICC	1:50 - 1:200

*Results are sample-specific. Please refer to your local assay conditions and test parameters for reference.

Overview

Description	Rabbit polyclonal antibody to SMAD3 (Phospho-S423)
Specificity	Recognizes endogenous levels of SMAD3 protein only when phosphorylated at S423.
Antibody Type	Primary antibody
Imnunogen	KLH-conjugated synthetic phosphopeptide corresponding to residues surrounding S423 of human SMAD3 protein. The exact sequence is proprietary.
Purification	The antibody was purified by immunogen affinity chromatography.
Molecular Weight	Predicted: 48 kD; Observed: 52 kD
Form/Buffer	Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.01% sodium azide.
Alternative Names	MADH3; Mothers against decapentaplegic homolog 3; MAD homolog 3; Mad3; Mothers against DPP homolog 3; hMAD-3; JV15-2; SMAD family member 3; SMAD 3; Smad3; hSMAD3
Gene Symbol	SMAD3
Entrez Gene	4088(Human); 17127(Mouse); 25631(Rat)
SwissProt	P84022(Human); Q8BUN5(Mouse); P84025(Rat)

*AREX continuously optimizes our products. Webpage content may not reflect the latest updates. For inquiries, please contact info@arexbio.com or your local distributor.
*Clone Number, Reactivity, Source/Host and Clonality can be found in the product name and Key Features section above.

Data

Western blot analysis of SMAD3 (Phospho-S423) expression in HeLa (A), Raw264.7 (B), H9C2 (C) whole cell lysates. (Predicted band size: 48 kD; Observed band size: 52 kD)

Immunofluorescent analysis of SMAD3 (Phospho-S423) staining in MPC5 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 °C in a hidified chamber. Cells were washed with PBST and incubated with a AREX® Fluor 488 -conjugated secondary antibody (green) in PBS at room temperature in the dark. Phalloidin - AREX® Fluor 594 was used to stain Actin filaments (red). DAPI was used to stain the cell nuclei (blue).

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Note

For Research Use Only. Not for use in diagnostic procedures.

FAQs

What are the main types of research antibodies and how do they differ?

Research antibodies are mainly divided into monoclonal antibodies and polyclonal antibodies. Monoclonal antibodies typically offer higher specificity and better batch-to-batch consistency, while polyclonal antibodies often provide stronger affinity but may show more variation between batches. The choice depends on your specific experimental needs.

How can I tell if a research antibody is suitable for my experiment?

It is recommended to carefully review the product datasheet for validated applications, species reactivity, recommended dilutions, and published references. For new antibodies, performing a small-scale validation with positive control samples is usually helpful.

Can improper storage of research antibodies affect experimental results?

Yes. Antibodies are sensitive to temperature, repeated freeze-thaw cycles, and contamination. Improper storage may lead to reduced activity, increased background, or weaker signals. It is best to follow the storage instructions provided in the product datasheet.

Why doesn’t the recommended dilution in the datasheet work well in my experiment?

The recommended dilution is based on the supplier’s test conditions. Factors such as sample type, fixation method, and detection system in your lab can influence the optimal working concentration. Performing a dilution series optimization in your own system is often necessary.

What precautions should I take when using a newly purchased research antibody for the first time?

It is advisable to briefly centrifuge the antibody (especially concentrated or lyophilized ones), then perform a small-scale pilot experiment using the recommended conditions. Recording the batch number and usage date is also helpful for future tracking.

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