EGFR (Phospho-Y869) Rabbit Polyclonal Antibody

Datasheet

MSDS

EGFR (Phospho-Y869) Rabbit Polyclonal Antibody

Key features and details

Rabbit polyclonal antibody to EGFR (Phospho-Y869)

Target: EGFR (Phospho-Y869)
Source/Host: Rabbit
Reactivity: Human
Clonality: Polyclonal
Applications: WB, IF/ICC
Conjugation: Unconjugated
Storage: at-20°C

Brand:

CAT.NO. : APA11518

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Size:

50μL 100μL

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Product Details

Background

Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses . Known ligands include EGF, TGFA/TGF-alpha, AREG, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF . Ligand binding triggers receptor homo- and/or heterodimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades. Activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STATs modules . May also activate the NF-kappa-B signaling cascade .

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 EGFR (Phospho-Y869)
Specificity	Recognizes endogenous levels of EGFR protein only when phosphorylated at Y869.
Antibody Type	Primary antibody
Imnunogen	KLH-conjugated synthetic phosphopeptide corresponding to residues surrounding Y869 of human EGFR protein. The exact sequence is proprietary.
Purification	The antibody was purified by immunogen affinity chromatography.
Molecular Weight	Predicted: 134 kD; Observed: 175 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	ERBB; ERBB1; HER1; Epidermal growth factor receptor; Proto-oncogene c-ErbB-1; Receptor tyrosine-protein kinase erbB-1
Gene Symbol	EGFR
Entrez Gene	1956(Human)
SwissProt	P00533(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 EGFR (Phospho-Y869) expression in Hela (A) whole cell lysates. (Predicted band size: 134 kD; Observed band size: 175 kD)

Immunofluorescent analysis of EGFR (Phospho-Y869) staining in A549 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 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 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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