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Purified anti-UPF1 Antibody

Pricing & Availability
Clone
W22273D (See other available formats)
Regulatory Status
RUO
Other Names
UPF1 RNA Helicase And ATPase, NORF1, PNORF1, Regulator Of Nonsense Transcripts 1, RENT1, Up-Frameshift Suppressor 1 Homolog, Nonsense MRNA Reducing Factor 1, UP Frameshift 1, Delta Helicase, HUpf1, UTF, Smg-2, Q92900
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
W22273D_Purified_anti-UPF1_Antibody_1_091624
Whole cell extracts (15 μg total protein per lane) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-UPF1 (clone W22273D) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • W22273D_Purified_anti-UPF1_Antibody_1_091624
    Whole cell extracts (15 μg total protein per lane) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-UPF1 (clone W22273D) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • W22273D_Purified_anti-UPF1_Antibody_2_091624
    A-431 cells were fixed with Fixation Buffer (Cat. No. 420801) for 10 minutes and permeabilized with 100% ice-cold methanol for 10 minutes, and blocked with 5% FBS at room temperature. Cells were then stained with Purified Rat IgG2a, κ Isotype Ctrl (Cat. No. 400502) (panel A) or Purified anti-UPF1 (clone W22273D) (panels B and C), followed by Alexa Fluor® 647 Goat anti-Rat IgG (Cat. No. 405416) (red). Nuclei were counterstained with DAPI (Cat. No. 422801) (blue). The images were captured in a Revvity Operetta CLS™ High Content Analysis System with a 63X objective and merged (panels A and C). Scale bar: 50 μm
  • W22273D_Purified_anti-UPF1_Antibody_3_091624
    IHC staining of Purified anti-UPF1 (clone W22273D) on formalin-fixed paraffin-embedded human testis tissue. Following antigen retrieval using 1X Tris-EDTA pH 9.0 Antigen Retrieval Buffer (Cat. No. 422704), the tissue was incubated with (panels A and C) and without (panels B and D) Purified anti-UPF1 (clone W22273D) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) (magenta) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (blue). Images were captured with 10X (panels A and B) and 40X objective (panels C and D) and merged. Scale bar: 50 μm
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667451 25 µg 144€
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667452 100 µg 356€
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Description

UPF1 is a crucial RNA helicase and an integral part of the nonsense-mediated mRNA decay (NMD) pathway, a cellular quality control mechanism that ensures the fidelity of gene expression by degrading mRNAs containing premature termination codons. UPF1 contains several key domains that contribute to its function: a N-terminal cysteine-histidine-rich domain (CH domain), a central helicase domain, and a C-terminal domain. The CH domain is involved in regulating the ATPase and helicase activities of UPF1, while the helicase domain facilitates ATP-dependent unwinding of RNA secondary structures. The C-terminal domain interacts with other NMD factors and regulatory proteins. 

UPF1 facilitates mRNA decay through two primary mechanisms: endonucleolytic cleavage and exonucleolytic decay. SMG6, an endonuclease, directly cleaves the mRNA near the PTC, while SMG5 and SMG7 recruit exonucleases to degrade the RNA from the 5’ and 3’ ends. UPF1's helicase activity is pivotal in remodeling RNA-protein complexes and unwinding secondary structures, thereby allowing exonucleases to efficiently degrade the target mRNA.

Beyond its role in NMD, UPF1 is involved in other RNA surveillance and regulatory pathways. It participates in Staufen-mediated mRNA decay (SMD), which targets specific mRNAs bound by the Staufen protein. UPF1 is also implicated in the regulation of histone mRNA decay and telomere maintenance, indicating its versatility in RNA metabolism.

Mutations or dysregulation of UPF1 and the NMD pathway have been linked to various human diseases, including cancer, neurodevelopmental disorders, and genetic diseases caused by nonsense mutations.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Recombinant fragment of human UPF1
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
IHC-P, ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.25 - 1.0 µg/mL. For immunohistochemistry, a concentration range of 1.0 - 10.0 µg/mL is suggested. For immunocytochemistry, a concentration range of  1.25 - 10.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

For use in immunohistochemistry on formalin-fixed paraffin-embedded tissue (IHC-P), it is recommended to perform antigen retrieval using Citrate Buffer, 10X (Cat. No. 420902) or Tris-EDTA pH 9.0 Antigen Retrieval Buffer (10X) (Cat. No. 422704).

For use in immunocytochemistry (ICC), it is recommended to fix/permeabilize with either of the following:
-Fixation buffer (Cat. No. 420801) with 0.5 % Triton X-100
-Fixation buffer (Cat. No. 420801) with 100% ice-cold methanol
-100% ice-cold methanol only

This antibody does not cross react with mouse UPF1.

Antigen Details

Structure
Contains a N-terminal cysteine-histidine-rich domain (CH domain), a central helicase domain, and a C-terminal domain
Distribution

Ubiquitously expressed in various tissues and cells. Found in the cytoplasm and nucleus

Function
Degrades mRNAs containing premature termination codons, preventing the production of truncated and potentially harmful proteins
Interaction
Participates in post-splicing messenger ribonucleoprotein (mRNP) and exon junction complex (EJC); Interacts with UPF2, UPF3A, UPF3B, SMG5, SMG7, and Staufen1
Molecular Family
Enzymes and Regulators
Antigen References
  1. Kim YK, et al. 2005. Cell. 120:195-208.
  2. Chamieh H, et al. 2008. Nat Struct Mol Biol. 15:85-93.
  3. Isken O, et al. 2008. Cell. 133:314-27.
  4. Hogg JR, et al. 2010. Cell. 143:379-89.
Gene ID
5976 View all products for this Gene ID
UniProt
View information about UPF1 on UniProt.org
Go To Top Version: 1    Revision Date: 09-16-2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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