Western Blot (WB)
(Figure 1. Western blot analysis of UPF3A using anti-UPF3A antibody (MBS1753789).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: human Hela whole cell lysatesLane 2: human HepG2 whole cell lysatesLane 3: human HEK293 whole cell lysatesLane 4: rat PC-12 whole cell lysatesLane 5: mouse NIH/3T3 whole cell 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-UPF3A antigen affinity purified polyclonal antibody (Catalog # MBS1753789) 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 UPF3A at approximately 51KD. The expected band size for UPF3A is at 51KD. )
Rabbit UPF3A Polyclonal Antibody | anti-UPF3A antibody
Anti-UPF3A Antibody
ICC/IF: 5ug/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 UPF3A using anti-UPF3A antibody (MBS1753789).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: human Hela whole cell lysatesLane 2: human HepG2 whole cell lysatesLane 3: human HEK293 whole cell lysatesLane 4: rat PC-12 whole cell lysatesLane 5: mouse NIH/3T3 whole cell 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-UPF3A antigen affinity purified polyclonal antibody (Catalog # MBS1753789) 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 UPF3A at approximately 51KD. The expected band size for UPF3A is at 51KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of UPF3A using anti-UPF3A antibody (MBS1753789).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: human Hela whole cell lysatesLane 2: human HepG2 whole cell lysatesLane 3: human HEK293 whole cell lysatesLane 4: rat PC-12 whole cell lysatesLane 5: mouse NIH/3T3 whole cell 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-UPF3A antigen affinity purified polyclonal antibody (Catalog # MBS1753789) 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 UPF3A at approximately 51KD. The expected band size for UPF3A is at 51KD. )
Immunofluorescence (IF)
(Figure 2. IF analysis of UPF3A using anti- UPF3A antibody (MBS1753789).UPF3A was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-UPF3A Antibody (MBS1753789) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Immunofluorescence (IF)
(Figure 2. IF analysis of UPF3A using anti- UPF3A antibody (MBS1753789).UPF3A was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-UPF3A Antibody (MBS1753789) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of HL-60 cells using anti-UPF3A antibody (MBS1753789).Overlay histogram showing HL-60 cells stained with MBS1753789 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-UPF3A Antibody (MBS1753789, 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 3. Flow Cytometry analysis of HL-60 cells using anti-UPF3A antibody (MBS1753789).Overlay histogram showing HL-60 cells stained with MBS1753789 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-UPF3A Antibody (MBS1753789, 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. Gross, M. B. Personal Communication. Baltimore, Md. 2/28/2011.
3. Kim, V. N., Kataoka, N., Dreyfuss, G. Role of the nonsense-mediated decay factor hUpf3 in the splicing-dependent exon-exon junction complex. Science 293: 1832-1836, 2001.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a protein that is part of a post-splicing multiprotein complex involved in both mRNA nuclear export and mRNA surveillance. The encoded protein is one of two functional homologs to yeast Upf3p. mRNA surveillance detects exported mRNAs with truncated open reading frames and initiates nonsense-mediated mRNA decay (NMD). When translation ends upstream from the last exon-exon junction, this triggers NMD to degrade mRNAs containing premature stop codons. This protein binds to the mRNA and remains bound after nuclear export, acting as a nucleocytoplasmic shuttling protein. It forms with Y14 a complex that binds specifically 20 nt upstream of exon-exon junctions. This gene is located on the long arm of chromosome 13. Two splice variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jul 2008]
Uniprot Description
UPF3A: Involved in nonsense-mediated decay (NMD) of mRNAs containing premature stop codons by associating with the nuclear exon junction complex (EJC) and serving as link between the EJC core and NMD machinery. Recruits UPF2 at the cytoplasmic side of the nuclear envelope and the subsequent formation of an UPF1-UPF2- UPF3 surveillance complex (including UPF1 bound to release factors at the stalled ribosome) is believed to activate NMD. However, UPF3A is shown to be only marginally active in NMD as compared to UPF3B. Binds spliced mRNA upstream of exon-exon junctions. In vitro, weakly stimulates translation. Belongs to the RENT3 family. 3 isoforms of the human protein are produced by alternative splicing.
Chromosomal Location of Human Ortholog: 13q34
Cellular Component: nucleoplasm; intracellular membrane-bound organelle; cytoplasm; plasma membrane; nucleus; cytosol
Molecular Function: protein binding; RNA binding; nucleotide binding; nucleocytoplasmic transporter activity
Biological Process: mRNA transport; positive regulation of translation; mRNA catabolic process, nonsense-mediated decay; gene expression; nucleocytoplasmic transport
Research Articles on UPF3A
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Product Notes
The UPF3A upf3a (Catalog #AAA1753789) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-UPF3A Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's UPF3A can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Human, Mouse, Rat| ICC/IF: 5ug/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 UPF3A upf3a 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 "UPF3A, 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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