FCM/FACS (Flow Cytometry)
(Figure 2. Flow Cytometry analysis of U937 cells using anti-OPA3 antibody (AAA127360).Overlay histogram showing U937 cells stained with AAA127360 (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-OPA3 Antibody (AAA127360, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)
Rabbit anti-Human OPA3 Polyclonal Antibody | anti-OPA3 antibody
Anti-OPA3 Antibody Picoband
Gene Names
OPA3; MGA3
Reactivity
Human
Applications
ELISA, Flow Cytometry, Functional Assay, Western Blot
Purity
Immunogen affinity purified.
Synonyms
OPA3, Antibody; Anti-OPA3 Antibody Picoband; AP-2 complex subunit mu; spindle and kinetochore associated complex subunit 2; Spindle and kinetochore-associated protein 2; Protein FAM33A; SKA2; FAM33A; anti-OPA3 antibody
Host
Rabbit
Reactivity
Human
Clonality
Polyclonal
Isotype
Rabbit IgG
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Applicable Applications for anti-OPA3 antibody
ELISA, FCM/FACS (Flow Cytometry), WB (Western Blot)
Immunogen
E coli-derived human OPA3 recombinant protein (Position: M1-K179).
Subcellular Localization
Secreted
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500ug/ml.
Cross Reactivity
No cross-reactivity with other proteins.
Protein Function
Component of the SKA1 complex, a microtubule-binding subcomplex of the outer kinetochore that is essential for proper chromosome segregation (PubMed:17093495, PubMed:19289083, PubMed:23085020). Required for timely anaphase onset during mitosis, when chromosomes undergo bipolar attachment on spindle microtubules leading to silencing of the spindle checkpoint (PubMed:17093495). The SKA1 complex is a component of the kinetochore-microtubule interface and ly associates with microtubules as oligomeric assemblies (PubMed:19289083). The complex facilitates the processive movement of microspheres along a microtubule in a depolymerization-coupled manner (PubMed:17093495, PubMed:19289083). In the complex, it is required for SKA1 localization (PubMed:19289083). Affinity for microtubules is synergistically enhanced in the presence of the ndc-80 complex and may allow the ndc-80 complex to track depolymerizing microtubules (PubMed:23085020).
Preparation and Storage
At -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months.Avoid repeated freezing and thawing.
FCM/FACS (Flow Cytometry)
(Figure 2. Flow Cytometry analysis of U937 cells using anti-OPA3 antibody (AAA127360).Overlay histogram showing U937 cells stained with AAA127360 (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-OPA3 Antibody (AAA127360, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)
FCM/FACS (Flow Cytometry)
(Figure 2. Flow Cytometry analysis of U937 cells using anti-OPA3 antibody (AAA127360).Overlay histogram showing U937 cells stained with AAA127360 (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-OPA3 Antibody (AAA127360, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)
WB (Western Blot)
(Figure 1. Western blot analysis of OPA3 using anti-OPA3 antibody (AAA127360).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 30 ug of sample under reducing conditions.Lane 1: human 293T whole cell lysates,Lane 2: human A549 whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-OPA3 antigen affinity purified polyclonal antibody (#AAA127360) at 0.5ug/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 with Tanon 5200 system. A specific band was detected for OPA3 at approximately 20 kDa. The expected band size for OPA3 is at 20 kDa.)
WB (Western Blot)
(Figure 1. Western blot analysis of OPA3 using anti-OPA3 antibody (AAA127360).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 30 ug of sample under reducing conditions.Lane 1: human 293T whole cell lysates,Lane 2: human A549 whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-OPA3 antigen affinity purified polyclonal antibody (#AAA127360) at 0.5ug/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 with Tanon 5200 system. A specific band was detected for OPA3 at approximately 20 kDa. The expected band size for OPA3 is at 20 kDa.)
Related Product Information for anti-OPA3 antibody
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.
Product Categories/Family for anti-OPA3 antibody
References
1. Anikster, Y., Kleta, R., Shaag, A., Gahl, W. A., Elpeleg, O. Type III 3-methylglutaconic aciduria (optic atrophy plus syndrome, or Costeff optic atrophy syndrome): identification of the OPA3 gene and its founder mutation in Iraqi Jews. Am. J. Hum. Genet. 69: 1218-1224, 2001. 2. Arif, B., Kumar, K. R., Seibler, P., Vulinovic, F., Fatima, A., Winkler, S., Nurnberg, G., Thiele, H., Nurnberg, P., Jamil, A. Z., Bruggemann, A., Abbas, G., Klein, C., Lohmann, K. A novel OPA3 mutation revealed by exome sequencing: an example of reverse phenotyping. JAMA Neurol. 70: 783-787, 2013. 3. Carmi, N., Lev, D., Leshinsky-Silver, E., Anikster, Y., Blumkin, L., Kivity, S., Lerman-Sagie, T., Zerem, A. Atypical presentation of Costeff syndrome--severe psychomotor involvement and electrical status epilepticus during slow wave sleep. Europ. J. Paediat. Neurol. 19: 733-736, 2015.
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
Molecular Weight
179
NCBI Official Full Name
Optic atrophy 3 protein
NCBI Official Synonym Full Names
optic atrophy 3 (autosomal recessive, with chorea and spastic paraplegia)
NCBI Official Symbol
OPA3
NCBI Official Synonym Symbols
MGA3
NCBI Protein Information
optic atrophy 3 protein; Optic atrophy 3 (Iraqi-Jewish 'optic atrophy plus')
UniProt Protein Name
Optic atrophy 3 protein
UniProt Gene Name
OPA3
UniProt Entry Name
OPA3_HUMAN
Similar Products
Product Notes
The OPA3 opa3 (Catalog #AAA127360) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-OPA3 Antibody Picoband reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's OPA3 can be used in a range of immunoassay formats including, but not limited to, ELISA, FCM/FACS (Flow Cytometry), WB (Western Blot). Researchers should empirically determine the suitability of the OPA3 opa3 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 "OPA3, 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
If you are ready to order, navigate to Shopping Cart and get ready to checkout.