EGFR Rabbit Monoclonal Antibody(ARA731)

Datasheet

MSDS

Key features and details

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

Brand:

CAT.NO. : ARA6518

US$ Please choose

Size:

100μL

Trail, Bulk size or Custom requests Please contact us

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	1:5000-1:10000
IHC	1:100-1:200
IF/ICC	1:200-1:800
FC	1:50-1:200
IP	1:10

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

Overview

Description	Rabbit Monoclonal Antibody to EGFR
Antibody Type	Primary antibody
Predicted MW	134kDa
Immunogen	A synthetic peptide corresponding to EGFR residues within the aa1000-1100 (intracellular) of human EGFR was used as an immunogen.
Purification	ProA affinity purified IgG
Form/Buffer	PBS 59%, Sodium azide 0.01%, Glycerol 40%, BSA 0.06%.
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); 13649(Mouse)
Swissprot	P00533

*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

All lanes: Anti-EGFR antibody at 1:5,000 dilution
Predicted MW: 134 kDa
Observed MW: 175 kDa

Lane 1: PC-3
Lane 2: SH-SY5Y
Lane 3: A549
Lane 4: Hela
Lane 5: A431
Lane 6: HaCat
Lane 7: Mu skin
Lane 8: Rat skin

Lysate at 10 μg per lane
2nd Ab:
G&R HRP(H+L) 1:10,000
Exposure: 100s

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung cancer tissue labelling EGFR with ARA731 at 1:200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0.

ARA731 staining EGFR in A431 cells by IF/ICC (immunofluorescence/immunocytochemistry). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% goat serum for half an hour at room temperature. Samples were incubated with primary antibody (1:200) at 4°C. An Alexa Fluor® 488-conjugated Goat Anti-Rabbit IgG polyclonal was used as the secondary antibody (1:500). DAPI (blue) was used as the nuclear counter stain.
Control: PBS and secondary antibody, An Alexa Fluor® 488-conjugated Goat Anti-Rabbit IgG (1:500).

Overlay histogram showing A431 cells stained with ARA731 (Red). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% TritonX-100 for 15 min. The cells were then incubated in the antibody (ARA731, 1:200 dilution) in 1x PBS/1% BSA for 30 min at room temperature. The secondary antibody used was a Goat Anti-Rabbit Alexa Fluor® 488 (IgG H+L) at 1:2,000 dilution for 20 min at room temperature. Unlabelled sample (Black) was used as a control.

EGFR was immunoprecipitated from 0.4mg of A431 whole cell lysate with ARA731 at 1:10 dilution.
2nd Ab:
GAR HRP for IP 1:500
Lane 1: ARA731 IP in A431 whole cell lysate
Lane 2: PBS instead of ARA731 in A431 whole cell lysate
Lane 3: A431 whole cell lysate, 10 μg (input)
Exposure: 120s

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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