HIF PHD3 Rabbit Polyclonal Antibody

MSDS

Key features and details

Rabbit polyclonal antibody to HIF PHD3

Target: HIF PHD3
Source/Host: Rabbit
Reactivity: Human, Mouse, Rat
Clonality: Polyclonal
Applications: WB, IF/ICC
Conjugation: Unconjugated
Storage: at-20°C

Brand:

CAT.NO. : APA15525

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Size:

50μL 100μL

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Product Details

Background

Prolyl hydroxylase that mediates hydroxylation of proline residues in target proteins, such as PKM, TELO2, ATF4, GPX4 and HIF1A . Target proteins are preferentially recognized via a LXXLAP motif. Cellular oxygen sensor that catalyzes, under normoxic conditions, the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins . Hydroxylates a specific proline found in each of the oxygen-dependent degradation (ODD) domains (N-terminal, NODD, and C-terminal, CODD) of HIF1A . Also hydroxylates HIF2A . Has a preference for the CODD site for both HIF1A and HIF2A . Hydroxylation on the NODD site by EGLN3 appears to require prior hydroxylation on the CODD site .

Application

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

WB	1:500 - 1:2000
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 HIF PHD3
Specificity	Recognizes endogenous levels of HIF PHD3 protein
Antibody Type	Primary antibody
Imnunogen	Recombinant fusion protein of human HIF PHD3. The exact sequence is proprietary.
Purification	The antibody was purified by immunogen affinity chromatography.
Molecular Weight	Predicted: 27 kD; Observed: 32 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	Egl nine homolog 3; HPH-1; Hypoxia-inducible factor prolyl hydroxylase 3; HIF-PH3; HIF-prolyl hydroxylase 3; HPH-3; Prolyl hydroxylase domain-containing protein 3; PHD3
Gene Symbol	EGLN3
Entrez Gene	112399(Human); 112407(Mouse); 54702(Rat)
SwissProt	Q9H6Z9(Human); Q91UZ4(Mouse); Q62630(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 HIF PHD3 expression in mouse liver (A) whole cell lysates. (Predicted band size: 27 kD; Observed band size: 32 kD)

Immunofluorescent analysis of HIF PHD3 staining in U2OS 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 Alexa Fluor 488-conjugated secondary antibody (green) 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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