FCM/FACS (Flow Cytometry)
(Figure 2. Flow Cytometry analysis of U251 cells using anti-Spt6/SUPT6H antibody (AAA127377).Overlay histogram showing U251 cells stained with AAA127377 (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-Spt6/SUPT6H Antibody (AAA127377, 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 Spt6/SUPT6H Polyclonal Antibody | anti-Spt6/SUPT6H antibody
Anti-Spt6/SUPT6H Antibody Picoband
Reactivity
Human, Monkey, Mouse, Rat
Applications
ELISA, Flow Cytometry, Functional Assay, Western Blot
Purity
Immunogen affinity purified.
Synonyms
Spt6/SUPT6H, Antibody; Anti-Spt6/SUPT6H Antibody Picoband; mediator complex subunit 4; Mediator of RNA polymerase II transcription subunit 4; Activator-recruited cofactor 36 kDa component; ARC36; Mediator complex subunit 4; TRAP/SMCC/PC2 subunit p36 subunit; Vitamin D3 receptor-interacting protein complex 36 kDa component; DRIP36; MED4; VDRIP; HSPC126; anti-Spt6/SUPT6H antibody
Host
Rabbit
Reactivity
Human, Monkey, Mouse, Rat
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-Spt6/SUPT6H antibody
ELISA, FCM/FACS (Flow Cytometry), WB (Western Blot)
Immunogen
E coli-derived human Spt6/SUPT6H recombinant protein (Position: A564-Q1639).
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 U251 cells using anti-Spt6/SUPT6H antibody (AAA127377).Overlay histogram showing U251 cells stained with AAA127377 (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-Spt6/SUPT6H Antibody (AAA127377, 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 U251 cells using anti-Spt6/SUPT6H antibody (AAA127377).Overlay histogram showing U251 cells stained with AAA127377 (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-Spt6/SUPT6H Antibody (AAA127377, 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 Spt6/SUPT6H using anti-Spt6/SUPT6H antibody (AAA127377).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 Hela whole cell lysates,Lane 2: human 293T whole cell lysates,Lane 3: monkey COS-7 whole cell lysates,Lane 4: rat testis tissue lysates,Lane 5: mouse NIH/3T3 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-Spt6/SUPT6H antigen affinity purified polyclonal antibody (#AAA127377) 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 Spt6/SUPT6H at approximately 260 kDa. The expected band size for Spt6/SUPT6H is at 199 kDa.)
WB (Western Blot)
(Figure 1. Western blot analysis of Spt6/SUPT6H using anti-Spt6/SUPT6H antibody (AAA127377).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 Hela whole cell lysates,Lane 2: human 293T whole cell lysates,Lane 3: monkey COS-7 whole cell lysates,Lane 4: rat testis tissue lysates,Lane 5: mouse NIH/3T3 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-Spt6/SUPT6H antigen affinity purified polyclonal antibody (#AAA127377) 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 Spt6/SUPT6H at approximately 260 kDa. The expected band size for Spt6/SUPT6H is at 199 kDa.)
Related Product Information for anti-Spt6/SUPT6H 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-Spt6/SUPT6H antibody
References
1. Andrulis, E. D., Werner, J., Nazarian, A., Erdjument-Bromage, H., Tempst, P., Lis, J. T. The RNA processing exosome is linked to elongating RNA polymerase II in Drosophila. Nature 420: 837-841, 2002.2. Chiang, P.-W., Wang, S., Smithivas, P., Song, W.-J., Ramamoorthy, S., Hillman, J., Puett, S., Van Keuren, M. L., Crombez, E., Kumar, A., Glover, T. W., Miller, D. E., Tsai, C.-H., Blackburn, C. C., Chen, X.-N., Sun, Z., Cheng, J.-F., Korenberg, J. R., Kurnit, D. M. Identification and analysis of the human and murine putative chromatin structure regulator SUPT6H and Supt6h. Genomics 34: 328-333, 1996. 3. Kaplan, C. D., Laprade, L., Winston, F. Transcription elongation factors repress transcription initiation from cryptic sites. Science 301: 1096-1099, 2003.
NCBI and Uniprot Product Information
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Product Notes
The Spt6/SUPT6H supt6h (Catalog #AAA127377) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-Spt6/SUPT6H Antibody Picoband reacts with Human, Monkey, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's Spt6/SUPT6H 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 Spt6/SUPT6H supt6h 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 "Spt6/SUPT6H, 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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