FCM/FACS (Flow Cytometry)
(Figure 2. Flow Cytometry analysis of SH-SY5Y cells using anti-RUFY3 antibody (AAA127584).Overlay histogram showing SH-SY5Y cells stained with AAA127584 (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-RUFY3 Antibody (AAA127584, 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 (Red line) was also used as a control.)
Rabbit RUFY3 Polyclonal Antibody | anti-RUFY3 antibody
Anti-RUFY3 Antibody Picoband
Gene Names
RUFY3; RIPX; SINGAR1; ZFYVE30
Reactivity
Human, Mouse, Rat
Applications
Flow Cytometry, Functional Assay, Western Blot, ELISA
Purity
Immunogen affinity purified.
Synonyms
RUFY3, Antibody; Anti-RUFY3 Antibody Picoband; CD72 molecule; Mediator of RNA polymerase II transcription subunit 20; Mediator complex subunit 20; TRF-proximal protein homolog; hTRFP; MED20; TRFP; anti-RUFY3 antibody
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Pre-B-cells and B-cells but not terminally differentiated plasma cells.
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-RUFY3 antibody
FCM/FACS (Flow Cytometry), WB (Western Blot), ELISA
Immunogen
E coli-derived human RUFY3 recombinant protein (Position: M1-H469).
Subcellular Localization
Nucleus
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 Mediator complex, a coactivator involved in the regulated transcription of nearly all RNA polymerase II-dependent genes. Mediator functions as a bridge to convey information from gene-specific regulatory proteins to the basal RNA polymerase II transcription machinery. Mediator is recruited to promoters by interactions with regulatory proteins and serves as a scaffold for the assembly of a functional preinitiation complex with RNA polymerase II and the general transcription factors.
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 SH-SY5Y cells using anti-RUFY3 antibody (AAA127584).Overlay histogram showing SH-SY5Y cells stained with AAA127584 (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-RUFY3 Antibody (AAA127584, 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 (Red line) was also used as a control.)
FCM/FACS (Flow Cytometry)
(Figure 2. Flow Cytometry analysis of SH-SY5Y cells using anti-RUFY3 antibody (AAA127584).Overlay histogram showing SH-SY5Y cells stained with AAA127584 (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-RUFY3 Antibody (AAA127584, 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 (Red line) was also used as a control.)
WB (Western Blot)
(Figure 1. Western blot analysis of RUFY3 using anti-RUFY3 antibody (AAA127584).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: rat brain tissue lysates,Lane 2: mouse brain tissue 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-RUFY3 antigen affinity purified polyclonal antibody (#AAA127584) 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 RUFY3 at approximately 53-65 kDa. The expected band size for RUFY3 is at 53-65 kDa.)
WB (Western Blot)
(Figure 1. Western blot analysis of RUFY3 using anti-RUFY3 antibody (AAA127584).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: rat brain tissue lysates,Lane 2: mouse brain tissue 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-RUFY3 antigen affinity purified polyclonal antibody (#AAA127584) 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 RUFY3 at approximately 53-65 kDa. The expected band size for RUFY3 is at 53-65 kDa.)
Related Product Information for anti-RUFY3 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-RUFY3 antibody
References
1. Hertz, N. T., Adams, E. L., Weber, R. A., Shen, R. J., O'Rourke, M. K., Simon, D. J., Zebroski, H., Olsen, O., Morgan, C. W., Mileur, T. R., Hitchcock, A. M., Sinnott Armstrong, N. A., Wainberg, M., Bassik, M. C., Molina, H., Wells, J. A., Tessier-Lavigne, M. Neuronally enriched RUFY3 is required for caspase-mediated axon degeneration. Neuron 103: 412-422, 2019. 2. Mori, T., Wada, T., Suzuki, T., Kubota, Y., Inagaki, N. Singar1, a novel RUN domain-containing protein, suppresses formation of surplus axons for neuronal polarity. J. Biol. Chem. 282: 19884-19893, 2007.3. Nagase, T., Ishikawa, K., Suyama, M., Kikuno, R., Hirosawa, M., Miyajima, N., Tanaka, A., Kotani, H., Nomura, N., Ohara, O. Prediction of the coding sequences of unidentified human genes. XII. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. DNA Res. 5: 355-364, 1998.
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
Molecular Weight
64,721 Da
NCBI Official Full Name
protein RUFY3 isoform 1
NCBI Official Synonym Full Names
RUN and FYVE domain containing 3
NCBI Official Symbol
RUFY3
NCBI Official Synonym Symbols
RIPX; SINGAR1; ZFYVE30
NCBI Protein Information
protein RUFY3; rap2 interacting protein x; single axon-regulated protein; single axon-related 1
UniProt Protein Name
Protein RUFY3
UniProt Gene Name
RUFY3
UniProt Synonym Gene Names
KIAA0871; RIPx; Singar
UniProt Entry Name
RUFY3_HUMAN
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Product Notes
The RUFY3 rufy3 (Catalog #AAA127584) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-RUFY3 Antibody Picoband reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's RUFY3 can be used in a range of immunoassay formats including, but not limited to, FCM/FACS (Flow Cytometry), WB (Western Blot), ELISA. Researchers should empirically determine the suitability of the RUFY3 rufy3 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 "RUFY3, 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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