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Product Info Summary
| SKU: | A04516-3 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Monkey, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-EYA4 Antibody Picoband®
SKU/Catalog Number
A04516-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-EYA4 Antibody Picoband® catalog # A04516-3. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Monkey, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-EYA4 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04516-3)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human EYA4 recombinant protein (Position: R28-E205).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A04516-3 is reactive to EYA4 in Human, Monkey, Rat
Observed Molecular Weight
50 kDa
Calculated molecular weight
69.5 kDa
Background of EYA4
This gene encodes a member of the eyes absent (EYA) family of proteins. The encoded protein may act as a transcriptional activator through its protein phosphatase activity, and it may be important for eye development, and for continued function of the mature organ of Corti. Mutations in this gene are associated with postlingual, progressive, autosomal dominant hearing loss at the deafness, autosomal dominant non-syndromic sensorineural 10 locus. The encoded protein is also a putative oncogene that mediates DNA repair, apoptosis, and innate immunity following DNA damage, cellular damage, and viral attack. Defects in this gene are also associated with dilated cardiomyopathy 1J. Alternative splicing results in multiple transcript variants encoding distinct isoforms.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A04516-3 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human, Monkey, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: monkey skeletal muscle tissue, human A431 whole cell, human HT1080 whole cell, rat skeletal muscle tissue
IHC: human mammary cancer tissue, human rectal cancer tissue
FCM: HELA cell
Validation Images & Assay Conditions
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Western blot analysis of EYA4 using anti-EYA4 antibody (A04516-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: monkey skeletal muscle tissue lysates,
Lane 2: human A431 whole cell lysates,
Lane 3: human HT1080 whole cell lysates,
Lane 4: rat skeletal muscle tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-EYA4 antigen affinity purified polyclonal antibody (Catalog # A04516-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for EYA4 at approximately 50KD. The expected band size for EYA4 is at 50KD.
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IHC analysis of EYA4 using anti-EYA4 antibody (A04516-3).
EYA4 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-EYA4 Antibody (A04516-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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IHC analysis of EYA4 using anti-EYA4 antibody (A04516-3).
EYA4 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-EYA4 Antibody (A04516-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Flow Cytometry analysis of HELA cells using anti-EYA4 antibody (A04516-3).
Overlay histogram showing HELA cells stained with A04516-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-EYA4 Antibody (A04516-3, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-EYA4 Antibody Picoband® (A04516-3)
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