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
Overview
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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
See all applications and species dataApplication Species ICC/IF HumanIP HumanWB Human -
Immunogen
Synthetic peptide within Human EGFR. The exact sequence is proprietary.
(Peptide available asab179559) -
Positive control
- A431 cells. IP: A431 cell lysate
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y39 -
Isotype
IgG -
Research areas
Images
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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 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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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.
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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 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.
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Immunofluorescent analysis of EGFR (phospho Y1086) expression in A431 cell culture, using 1/100 ab32086.
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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?
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