Antibodies, Proteins, Kits and Reagents for Life Science

304 886 ₸ Product size

100 µl 304 886 ₸

Order now and get it on Tuesday March 09, 2021

Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [Y39] to EGFR (phospho Y1086)
Suitable for: WB, IP, Dot blot, ICC/IF
Reacts with: Human

Product name

Anti-EGFR (phospho Y1086) antibody [Y39]
See all EGFR primary antibodies
Description

Rabbit monoclonal [Y39] to EGFR (phospho Y1086)
Host species

Rabbit
Specificity

This antibody only detects EGFR phosphorylated on Tyrosine 1086 of the mature human isoform 1.

Tested Applications & Species

Application	Species
ICC/IF	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human EGFR. The exact sequence is proprietary.
(Peptide available as ab179559)
Positive control
- A431 cells. IP: A431 cell lysate
General notes
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

Y39
Isotype

IgG
Research areas

Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)

All lanes : Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086) at 1/3000 dilution

Lane 1 : Untreated A431 (Human epidermoid carcinoma cell line) whole cell lysates
Lane 2 : A431 (Human epidermoid carcinoma cell line) treated with 100 ng/ml EGF for 10 minutes whole cell lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 134 kDa
Observed band size: 170 kDa
why is the actual band size different from the predicted?

Exposure time: 10 seconds

Blocking and dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)

Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma cell line) cells labeling EGFR (phospho Y1086) with ab32086 at 1/100, 3 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077, a AlexaFluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1000, 2 μg/ml. Cells were counterstained with ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200, 2.5 μg/ml. Nuclear stain was DAPI (blue).

The green staining on the membrane was increased in the EGF (100ng/ml, 10min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signaling was obviously decreased.

For the pan antibody, there was no great difference after EGF (100ng/ml, 10min) or EGF (100ng/ml, 10min) + LP treatment. The data showed mostly membranous staining.
Immunoprecipitation - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)

Purified ab32086 at 1/20 dilution (1µg) immunoprecipitating EGFR in A431 treated with 100ng/mL EGF for 30min whole cell lysate.
Lane 1 (input): A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/mL EGF for 30min whole cell lysate 10µg
Lane 2 (+): ab32086 + A431 treated with 100ng/mL EGF for 30min whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32086 in A431 treated with 100ng/mL EGF for 30min whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 175 kDa
Dot Blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)

Dot blot analysis of EGFR (phospho Y1086) phospho peptide (Lane 1), EGFR non-phospho peptide (Lane 2), labeling EGFR (phospho Y1086) with ab32068 at a dilution of 1/1000. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/100000.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 5 seconds.
Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)

Immunofluorescent analysis of EGFR (phospho Y1086) expression in A431 cell culture, using 1/100 ab32086.
Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)

All lanes : Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086) at 1/10000 dilution

Lane 1 : Untreated A431 cell lysate
Lane 2 : A431 cell lysate treated with EGF

Predicted band size: 134 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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