Name: Anti-RENT1/hUPF1 antibody [EPR4681]
SKU: ab109363
Rating: 5 (2 reviews)

Rabbit Recombinant Monoclonal RENT1/hUPF1 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse samples. Cited in 25 publications.

Images

Immunoprecipitation - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Expected	Expected	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse. Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10000 - 1/50000	Notes -
Species Mouse	Dilution info 1/10000 - 1/50000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes For unpurified use at 1/100 - 1/250.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/100	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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Target data

See full target information UPF1

Function

RNA-dependent helicase required for nonsense-mediated decay (NMD) of aberrant mRNAs containing premature stop codons and modulates the expression level of normal mRNAs (PubMed:11163187, PubMed:16086026, PubMed:18172165, PubMed:21145460, PubMed:21419344, PubMed:24726324). Is recruited to mRNAs upon translation termination and undergoes a cycle of phosphorylation and dephosphorylation; its phosphorylation appears to be a key step in NMD (PubMed:11544179, PubMed:25220460). Recruited by release factors to stalled ribosomes together with the SMG1C protein kinase complex to form the transient SURF (SMG1-UPF1-eRF1-eRF3) complex (PubMed:19417104). In EJC-dependent NMD, the SURF complex associates with the exon junction complex (EJC) (located 50-55 or more nucleotides downstream from the termination codon) through UPF2 and allows the formation of an UPF1-UPF2-UPF3 surveillance complex which is believed to activate NMD (PubMed:21419344). Phosphorylated UPF1 is recognized by EST1B/SMG5, SMG6 and SMG7 which are thought to provide a link to the mRNA degradation machinery involving exonucleolytic and endonucleolytic pathways, and to serve as adapters to protein phosphatase 2A (PP2A), thereby triggering UPF1 dephosphorylation and allowing the recycling of NMD factors (PubMed:12554878). UPF1 can also activate NMD without UPF2 or UPF3, and in the absence of the NMD-enhancing downstream EJC indicative for alternative NMD pathways (PubMed:18447585). Plays a role in replication-dependent histone mRNA degradation at the end of phase S; the function is independent of UPF2 (PubMed:16086026, PubMed:18172165). For the recognition of premature termination codons (PTC) and initiation of NMD a competitive interaction between UPF1 and PABPC1 with the ribosome-bound release factors is proposed (PubMed:18447585, PubMed:25220460). The ATPase activity of UPF1 is required for disassembly of mRNPs undergoing NMD (PubMed:21145460). Together with UPF2 and dependent on TDRD6, mediates the degradation of mRNA harboring long 3'UTR by inducing the NMD machinery (By similarity). Also capable of unwinding double-stranded DNA and translocating on single-stranded DNA (PubMed:30218034).

Alternative names

KIAA0221, RENT1, UPF1, Regulator of nonsense transcripts 1, ATP-dependent helicase RENT1, Nonsense mRNA reducing factor 1, Up-frameshift suppressor 1 homolog, NORF1, hUpf1

Recommended products

Rabbit Recombinant Monoclonal RENT1/hUPF1 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse samples. Cited in 25 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR4681
Purification technique: Affinity purification Protein A
Specificity: The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Species reactivity
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species.
Please contact us for more information.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

RENT1/hUPF1 also known as regulator of nonsense transcripts 1 or human UPF1 is an important factor in RNA surveillance mechanisms. It is an ATP-dependent helicase with a molecular mass of approximately 130 kDa. This protein is ubiquitously expressed in various tissues highlighting its importance across different biological systems. Mechanically UPF1 plays an essential role in the nonsense-mediated mRNA decay (NMD) pathway where it interacts with other core NMD factors to identify and degrade mRNAs containing premature stop codons.

Biological function summary

RENT1/hUPF1 ensures the quality control of mRNA by facilitating the rapid degradation of defective transcripts and thereby preventing the synthesis of potentially harmful truncated proteins. As part of the NMD complex UPF1 partners with proteins like UPF2 and UPF3 to form a surveillance mechanism that maintains cellular homeostasis. The complex cooperates to distinguish transcripts that require elimination from those necessary for normal cellular function.

Pathways

RENT1/hUPF1 integrates into essential cellular pathways beyond just NMD. The protein also links to the mRNA decay and translation initiation pathways playing significant roles alongside proteins such as SMG1 and eRF1. In these pathways UPF1 regulates gene expression by modulating mRNA stability and protein synthesis ensuring that only correctly processed mRNAs translate into proteins.

Associated diseases and disorders

RENT1/hUPF1 connects to conditions like neurodevelopmental disorders and cancer. Dysregulation of UPF1 activity can lead to improper mRNA decay contributing to the accumulation of faulty transcripts which may drive disease pathogenesis. In particular UPF1 alterations are linked to the disruption of pathways involving key proteins like p53 which are critical in tumor suppression. Understanding UPF1's role in these disorders could present opportunities for therapeutic interventions.

Product promise

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12 product images

Immunoprecipitation - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
ab109363 (purified) at 1:30 dilution (2μg) immunoprecipitating RENT1/hUPF1 in Raji whole cell lysate.
Lane 1 (input): Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10μg
Lane 2 (+): ab109363 & Raji whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab109363 in Raji whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
Predicted band size: 124 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling RENT1/hUPF1 with Purified ab109363 at 1:100 dilution (5.14 μg/ml). Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunocytochemistry/ Immunofluorescence - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RENT1/hUPF1 with Purified ab109363 at 1:100 dilution (5.1 μg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Flow Cytometry (Intracellular) - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RENT1/hUPF1 with Purified ab109363 at 1/50 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Western blot - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
All lanes: Western blot - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363) at 1/10000 dilution
Lane 1: HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysates at 20 µg
Lane 2: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 124 kDa
Observed band size: 130 kDa
Western blot - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
This image was generated using the unpurified version of the product.
All lanes: Western blot - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363) at 1/10000 dilution
Lane 1: HuT-78 cell lysate at 10 µg
Lane 2: Raji cell lysate at 10 µg
Lane 3: SH-SY5Y cell lysate at 10 µg
Lane 4: HeLa cell lysate at 10 µg
Performed under reducing conditions.
Predicted band size: 124 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
ab109363, at 1/100, staining RENT1/hUPF1 in Human kidney tissue by immunohistochemistry.
This image was generated using the unpurified version of the product.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling RENT1/hUPF1 with purified ab109363 at 1/500. Cells were fixed with 100% methanol. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).
Secondary Only Control: PBS was used instead of the primary antibody as the negative control
This image was generated using the unpurified version of the product.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
ab109363, at 1/100, staining RENT1/hUPF1 in Human transitional cell carcinoma tissue by immunohistochemistry.
This image was generated using the unpurified version of the product.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
ab109363, at 1/100, staining RENT1/hUPF1 in HeLa cells by immunofluorescence.
This image was generated using the unpurified version of the product.
Flow Cytometry (Intracellular) - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
Overlay histogram showing HeLa cells stained with ab109363 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109363, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This image was generated using the unpurified version of the product.
Image collected and cropped by CiteAb under a CC-BY license from the publication
Western blot - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363)
RENT1/hUPF1 western blot using anti-RENT1/hUPF1 antibody [EPR4681] ab109363. Publication image and figure legend from Mooney, C. M., Jiménez-Mateos, E. M., et al., 2017, Sci Rep, PubMed 28128343.

ab109363 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab109363 please see the product overview.
NMD proteins are increased after SE in mice and Upf1 is increased in human TLE samples.(a–d) Protein levels of Upf1, phosho-Upf1 (p-Upf1), Upf2 and Upf3b in the ipsilateral hippocampus in control (C) mice and at 1, 4, 8 and 24 hours (h) after SE were analysed by western blot and semi-quantified. (a) Upf1 levels significantly increased 8 and 24 h after SE (n = 4/group; ANOVA, Dunnett’s posthoc test ***p < 0.001, **p < 0.01). (b) p-Upf1 levels significantly increased 8 and 24 h after SE (n = 5/group; ANOVA, Dunnett’s posthoc test *p < 0.05 compared to control. (c) Upf2 levels were increased 8 h after SE (n = 4/group; ANOVA, Dunnett’s posthoc test *p < 0.05 compared to control). (d) Upf3b levels did not change after SE (n = 4/group; ANOVA, Dunnett’s posthoc test p = 0.88). (e) Upf1 protein levels were significantly higher in TLE patients with hippocampal sclerosis (HS) compared to post-mortem controls and TLE patients without HS (Controls n = 6; TLE without HS n = 3, TLE with HS n = 3; ANOVA with Bonferroni post-hoc test comparing all columns; p = 0.0072). Representative blots have been cropped to reduce unnecessary area.