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Rabbit Recombinant Monoclonal PUS1 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

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Images

Western blot - Anti-PUS1 antibody [EPR20181] (AB203010), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-PUS1 antibody [EPR20181] (AB203010), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-PUS1 antibody [EPR20181] (AB203010), expandable thumbnail
  • Western blot - Anti-PUS1 antibody [EPR20181] (AB203010), expandable thumbnail
  • Western blot - Anti-PUS1 antibody [EPR20181] (AB203010), 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
IPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Expected
Rat
Expected
Tested
Expected
Expected

Tested
Tested

Species
Human
Dilution info
1/30
Notes

-

Expected
Expected

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

-

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Rat
Dilution info
1/1000
Notes

-

Species
Human
Dilution info
1/1000
Notes

-

Tested
Tested

Species
Human
Dilution info
1/500
Notes

This antibody was not successful when we used it on RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells in ICC application. This antibody was not tested on rat cells in ICC.

Expected
Expected

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

-

Tested
Tested

Species
Human
Dilution info
1/400
Notes

-

Expected
Expected

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

-

Target data

Function

Pseudouridylate synthase that catalyzes pseudouridylation of tRNAs and mRNAs (PubMed:15772074, PubMed:24722331). Acts on positions 27/28 in the anticodon stem and also positions 34 and 36 in the anticodon of an intron containing tRNA (PubMed:24722331). Also catalyzes pseudouridylation of mRNAs: mediates pseudouridylation of mRNAs with the consensus sequence 5'-UGUAG-3' (PubMed:31477916, PubMed:35051350). Acts as a regulator of pre-mRNA splicing by mediating pseudouridylation of pre-mRNAs at locations associated with alternatively spliced regions (PubMed:35051350). Pseudouridylation of pre-mRNAs near splice sites directly regulates mRNA splicing and mRNA 3'-end processing (PubMed:35051350). Involved in regulation of nuclear receptor activity through pseudouridylation of SRA1 mRNA (PubMed:24722331).

Alternative names

PP8985, PUS1, Pseudouridylate synthase 1 homolog, tRNA pseudouridine synthase 1, tRNA pseudouridine(38-40) synthase, tRNA pseudouridylate synthase I, tRNA-uridine isomerase I

Recommended products

Rabbit Recombinant Monoclonal PUS1 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR20181
Purification technique
Affinity purification Protein A
Concentration
Loading...

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
Avoid freeze / thaw cycle

Notes

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

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.

Supplementary info

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

PUS1 also known as pseudouridine synthase 1 is an enzyme responsible for catalyzing the isomerization of uridine to pseudouridine in RNA molecules. This protein has a mass of approximately 50 kDa and expresses widely in both the cytoplasm and mitochondria of human cells. Pseudouridine is the most abundant RNA modification and plays an important role in RNA stability and structure. The presence of PUS1 can vary among cell types supporting its necessity in diverse cellular contexts.

Biological function summary

The modification of RNA by PUS1 improves the stability and folding of the RNA structure aiding its functionality. The enzyme is not part of a larger complex but acts independently to achieve its role in RNA modification. Pseudouridine created by PUS1 permits enhanced base stacking and hydrogen bonding which influences the translation process and the overall efficiency of protein synthesis. This modification is important for the proper functioning of tRNA and rRNA ensuring fidelity during protein translation.

Pathways

PUS1 plays an essential role in the RNA processing and modification pathways. Its activity integrates into pathways such as ribosome biogenesis and RNA quality control. Particularly it interacts with other RNA-binding proteins and enzymes like TRUB1 another pseudouridine synthase which also contributes to the maturation of structured RNAs. These pathways are vital for maintaining cellular homeostasis and responding to various physiological demands.

Associated diseases and disorders

Mutations in PUS1 are linked to mitochondrial myopathy and sideroblastic anemia a rare condition characterized by muscle weakness and anemia. These disorders illustrate the critical impact of RNA modification defects on cellular energy production and red blood cell formation. In these conditions PUS1 may interact with defective proteins leading to impaired pseudouridylation disrupting normal cellular functions and contributing to disease pathology. Understanding the role of PUS1 might provide insights into potential therapeutic strategies for such disorders.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Western blot - Anti-PUS1 antibody [EPR20181] (ab203010), expandable thumbnail

    Western blot - Anti-PUS1 antibody [EPR20181] (ab203010)

    Lanes 1- 4: Merged signal (red and green). Green - ab203010 observed at 45 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) observed at 50 kDa.

    ab203010 was shown to react with PUS1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human PUS1 knockout HEK-293T cell line ab266091 (knockout cell lysate Human PUS1 knockout HEK-293T cell lysate ab258158) was used. Wild-type HEK-293T and PUS1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab203010 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-PUS1 antibody [EPR20181] (ab203010) at 1/1000 dilution

    Lane 1: Wild-type HEK-293T cell lysate at 20 µg

    Lane 2: PUS1 knockout HEK-293T cell lysate at 20 µg

    Lane 2: Western blot - Human PUS1 knockout HEK-293T cell line (Human PUS1 knockout HEK-293T cell line ab266091)

    Lane 3: HeLa cell lysate at 20 µg

    Lane 4: Daudi cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 47 kDa

    Observed band size: 45 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-PUS1 antibody [EPR20181] (ab203010), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-PUS1 antibody [EPR20181] (ab203010)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PUS1 with ab203010 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cell line.

    The nuclear counterstain is DAPI (blue). Tubulin is detected with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.

    This antibody was not successful when we used it on RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells in ICC application. This antibody was not tested on rat cells in ICC.

  • Immunocytochemistry/ Immunofluorescence - Anti-PUS1 antibody [EPR20181] (ab203010), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-PUS1 antibody [EPR20181] (ab203010)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PUS1 with ab203010 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mitochondrial staining on HeLa cell line.

    The nuclear counterstain is DAPI (blue). COXIV is detected with Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker ab33985 (Anti-COX IV(mouse mAb)) at 1/1000 dilution followed by Goat anti-mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab203010 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.

    -ve control 2: Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker ab33985 (anti-COX IV(mouse mAb)) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Western blot - Anti-PUS1 antibody [EPR20181] (ab203010), expandable thumbnail

    Western blot - Anti-PUS1 antibody [EPR20181] (ab203010)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-PUS1 antibody [EPR20181] (ab203010) at 1/1000 dilution

    All lanes: Human skeletal muscle lysate at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 47 kDa

    Observed band size: 47 kDa

    Exposure time: 3min

  • Western blot - Anti-PUS1 antibody [EPR20181] (ab203010), expandable thumbnail

    Western blot - Anti-PUS1 antibody [EPR20181] (ab203010)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in UniProt.

    All lanes: Western blot - Anti-PUS1 antibody [EPR20181] (ab203010) at 1/5000 dilution

    Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) mitochondria lysate at 20 µg

    Lane 2: HeLa cytoplasm fraction lysate at 20 µg

    Lane 3: A431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 47 kDa

    Observed band size: 47 kDa

    Exposure time: 8s

  • Western blot - Anti-PUS1 antibody [EPR20181] (ab203010), expandable thumbnail

    Western blot - Anti-PUS1 antibody [EPR20181] (ab203010)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Based on sequence alignment, the antibody can recognize 2 isoforms, the predicted MW are 47kDa and 44kDa, respectively [PMID: 17056637].

    All lanes: Western blot - Anti-PUS1 antibody [EPR20181] (ab203010) at 1/1000 dilution

    All lanes: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 47 kDa

    Observed band size: 44 kDa, 47 kDa

    Exposure time: 3s

  • Immunoprecipitation - Anti-PUS1 antibody [EPR20181] (ab203010), expandable thumbnail

    Immunoprecipitation - Anti-PUS1 antibody [EPR20181] (ab203010)

    PUS1 was immunoprecipitated from 0.35 mg of HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate with ab203010 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab203010 at 1/1000 dilution. VeriBlot for IP Detection Reaction (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HEK-293 whole cell lysate, 10μg (Input).

    Lane 2: ab203010 IP in HEK-293 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab203010 in HEK-293 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 5 seconds.

    All lanes: Immunoprecipitation - Anti-PUS1 antibody [EPR20181] (ab203010)

    Predicted band size: 47 kDa

    Observed band size: 44 kDa, 47 kDa

  • Western blot - Anti-PUS1 antibody [EPR20181] (ab203010), expandable thumbnail

    Western blot - Anti-PUS1 antibody [EPR20181] (ab203010)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 and 2: 10 seconds; Lane 3: 3minutes; Lane 4,5,6 and 7: 10 seconds.

    All lanes: Western blot - Anti-PUS1 antibody [EPR20181] (ab203010) at 1/2000 dilution

    Lane 1: Mouse heart tissue lysate at 10 µg

    Lane 2: Mouse spleen tissue lysate at 10 µg

    Lane 3: Rat spleen tissue lysate at 10 µg

    Lane 4: C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

    Lane 5: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

    Lane 6: PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

    Lane 7: NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 47 kDa

    Observed band size: 47 kDa

  • Flow Cytometry (Intracellular) - Anti-PUS1 antibody [EPR20181] (ab203010), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-PUS1 antibody [EPR20181] (ab203010)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PUS1 with ab203010 at 1/400 dilution (red) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution was used as the secondary antibody.

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Product protocols

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