Western Blot (WB)
(Figure 1. Western blot analysis of EYA4 using anti-EYA4 antibody (MBS1753661).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 lysatesLane 2: human A431 whole cell lysatesLane 3: human HT1080 whole cell lysatesLane 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 # MBS1753661) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) with Tanon 5200 system. A specific band was detected for EYA4 at approximately 50KD. The expected band size for EYA4 is at 50KD. )
Rabbit EYA4 Polyclonal Antibody | anti-EYA4 antibody
Anti-EYA4 Antibody
IHC-P: 0.5-1ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of EYA4 using anti-EYA4 antibody (MBS1753661).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 lysatesLane 2: human A431 whole cell lysatesLane 3: human HT1080 whole cell lysatesLane 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 # MBS1753661) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) with Tanon 5200 system. A specific band was detected for EYA4 at approximately 50KD. The expected band size for EYA4 is at 50KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of EYA4 using anti-EYA4 antibody (MBS1753661).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 lysatesLane 2: human A431 whole cell lysatesLane 3: human HT1080 whole cell lysatesLane 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 # MBS1753661) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) with Tanon 5200 system. A specific band was detected for EYA4 at approximately 50KD. The expected band size for EYA4 is at 50KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of EYA4 using anti-EYA4 antibody (MBS1753661).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 (MBS1753661) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of EYA4 using anti-EYA4 antibody (MBS1753661).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 (MBS1753661) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of EYA4 using anti-EYA4 antibody (MBS1753661).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 (MBS1753661) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of EYA4 using anti-EYA4 antibody (MBS1753661).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 (MBS1753661) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Flow Cytometry (FC/FACS)
(Figure 4. Flow Cytometry analysis of HELA cells using anti-EYA4 antibody (MBS1753661).Overlay histogram showing HELA cells stained with MBS1753661 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-EYA4 Antibody (MBS1753661, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 4. Flow Cytometry analysis of HELA cells using anti-EYA4 antibody (MBS1753661).Overlay histogram showing HELA cells stained with MBS1753661 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-EYA4 Antibody (MBS1753661, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Depreux, F. F. S., Darrow, K., Conner, D. A., Eavey, R. D., Liberman, M. C., Seidman, C. E., Seidman, J. G. Eya4-deficient mice are a model for heritable otitis media. J. Clin. Invest. 118: 651-658, 2008.
3. Hildebrand, M. S., Coman, D., Yang, T., Gardner, R. J. M., Rose, E., Smith, R. J. H., Bahlo, M., Dahl, H. -H. M. A novel splice site mutation in EYA4 causes DFNA10 hearing loss. Am. J. Med. Genet. 143A: 1599-1604, 2007. Note: Erratum: Am. J. Med. Genet. 146A: 1099 only, 2008.
NCBI and Uniprot Product Information
NCBI Description
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. [provided by RefSeq, Jul 2014]
Uniprot Description
EYA4: Tyrosine phosphatase that specifically dephosphorylates 'Tyr-142' of histone H2AX (H2AXY142ph). 'Tyr-142' phosphorylation of histone H2AX plays a central role in DNA repair and acts as a mark that distinguishes between apoptotic and repair responses to genotoxic stress. Promotes efficient DNA repair by dephosphorylating H2AX, promoting the recruitment of DNA repair complexes containing MDC1. Its function as histone phosphatase probably explains its role in transcription regulation during organogenesis. May be involved in development of the eye. Defects in EYA4 are the cause of deafness autosomal dominant type 10 (DFNA10). DFNA10 is a form of sensorineural hearing loss. Sensorineural deafness results from damage to the neural receptors of the inner ear, the nerve pathways to the brain, or the area of the brain that receives sound information. Defects in EYA4 are the cause of cardiomyopathy dilated type 1J (CMD1J). Dilated cardiomyopathy is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death. Belongs to the HAD-like hydrolase superfamily. EYA family. 5 isoforms of the human protein are produced by alternative splicing.
Protein type: Protein phosphatase, tyrosine (non-receptor); EC 3.1.3.48
Chromosomal Location of Human Ortholog: 6q23
Cellular Component: cytoplasm; nucleus
Molecular Function: protein binding; metal ion binding; protein tyrosine phosphatase activity
Biological Process: anatomical structure morphogenesis; sensory perception of sound; transcription, DNA-dependent; regulation of transcription, DNA-dependent; visual perception; multicellular organismal development; chromatin modification; DNA repair
Disease: Cardiomyopathy, Dilated, 1j; Deafness, Autosomal Dominant 10
Research Articles on EYA4
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Product Notes
The EYA4 eya4 (Catalog #AAA1753661) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-EYA4 Antibody reacts with Human, Monkey, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's EYA4 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Human, Monkey, Rat| IHC-P: 0.5-1ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the EYA4 eya4 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "EYA4, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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