FOXO3 (Phospho-S253) Rabbit Polyclonal Antibody

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FOXO3 (Phospho-S253) Rabbit Polyclonal Antibody

Key features and details

Rabbit polyclonal antibody to FOXO3 (Phospho-S253)

Target: FOXO3 (Phospho-S253)
Source/Host: Rabbit
Reactivity: Human, Mouse, Rat
Clonality: Polyclonal
Applications: WB, IHC, IF/ICC
Conjugation: Unconjugated
Storage: at-20°C

Brand:

CAT.NO. : APA10121

US$ Please choose

Size:

50μL 100μL

Trail, Bulk size or Custom requests Please contact us

Product Details

Background

Transcriptional activator that recognizes and binds to the DNA sequence 5'-[AG]TAAA[TC]A-3' and regulates different processes, such as apoptosis and autophagy . Acts as a positive regulator of autophagy in skeletal muscle: in starved cells, enters the nucleus following dephosphorylation and binds the promoters of autophagy genes, such as GABARAP1L, MAP1LC3B and ATG12, thereby activating their expression, resulting in proteolysis of skeletal muscle proteins . Triggers apoptosis in the absence of survival factors, including neuronal cell death upon oxidative stress .

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
IHC	1:50 - 1:100
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 FOXO3 (Phospho-S253)
Specificity	Recognizes endogenous levels of FOXO3 protein only when phosphorylated at S253.
Antibody Type	Primary antibody
Imnunogen	KLH-conjugated synthetic phosphopeptide corresponding to residues surrounding S253 of human FOXO3 protein. The exact sequence is proprietary.
Purification	The antibody was purified by immunogen affinity chromatography.
Molecular Weight	Predicted: 71 kD; Observed: 97 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	FKHRL1; FOXO3A; Forkhead box protein O3; AF6q21 protein; Forkhead in rhabdomyosarcoma-like 1
Gene Symbol	FOXO3
Entrez Gene	2309(Human); 56484(Mouse)
SwissProt	O43524(Human); Q9WVH4(Mouse)

*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 FOXO3 (Phospho-S253) expression in HCT116 (A), Raw264.7 (B) whole cell lysates. (Predicted band size: 71 kD; Observed band size: 97 kD)

Immunohistochemical analysis of FOXO3 (Phospho-S253) staining in human prostate cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of FOXO3 (Phospho-S253) 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 DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark.

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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