SMAD5 Mouse Monoclonal Antibody(C3303)

Datasheet

MSDS

Key features and details

Mouse monoclonal antibody to SMAD5

Target: SMAD5
Source/Host: Mouse
Reactivity: Human
Clonality: Monoclonal
Applications: WB, IF/ICC, FC
Conjugation: Unconjugated
Storage: at-20°C

Brand:

CAT.NO. : AMA02915

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Size:

50μL 100μL

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Product Details

Background

Transcriptional regulator that plays a role in various cellular processes including embryonic development, cell differentiation, angiogenesis and tissue homeostasis . Upon BMP ligand binding to their receptors at the cell surface, is phosphorylated by activated type I BMP receptors (BMPRIs) and associates with SMAD4 to form a heteromeric complex which translocates into the nucleus acting as transcription factor . In turn, the hetero-trimeric complex recognizes cis-regulatory elements containing Smad Binding Elements (SBEs) to modulate the outcome of the signaling network . Non-phosphorylated SMAD5 has a cytoplasmic role in energy metabolism regulation by promoting mitochondrial respiration and glycolysis in response to cytoplasmic pH changes .

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 - 1:100
FC	1:50 - 1:100

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

Overview

Description	Mouse monoclonal antibody to SMAD5
Specificity	Recognizes endogenous levels of SMAD5 protein.
Antibody Type	Primary antibody
Imnunogen	Purified recombinant human SMAD5 (C-terminus) protein fragments expressed in E.coli.
Purification	The antibody was purified by immunogen affinity chromatography.
Molecular Weight	Predicted: 52 kD; Observed: 60 kD
Form/Buffer	Liquid in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide, pH 7.3.
Alternative Names	MADH5; Mothers against decapentaplegic homolog 5; MAD homolog 5; Mothers against DPP homolog 5; JV5-1; SMAD family member 5; SMAD 5; Smad5; hSmad5
Gene Symbol	SMAD5
Entrez Gene	4090(Human)
SwissProt	Q99717(Human)

*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 SMAD5 expression in Hela (A), Jurkat (B), K562 (C) whole cell lysates. (Predicted band size: 52 kD; Observed band size: 60 kD)

Immunofluorescent analysis of SMAD5 staining in HeLa 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.

Flow cytometric analysis of SMAD5 in Jurkat cells (red). Black line histogram represents the isotype control.

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 diagnostic, therapeutics, prophylactic or in vivo use.

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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