Pseudouridine Monoclonal antibody

Pseudouridine Monoclonal Antibody for Dot Blot, ELISA

Host / Isotype

Mouse / IgG1

Reactivity

chemical compound

Applications

Dot Blot, ELISA

Conjugate

Unconjugated

CloneNo.

1G7C7

Cat no : 68578-1-Ig

Synonyms

Ψ, 5-ribosyluracil

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Validation Data Gallery

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  ELISA experiment of Pseudouridine using 68578-1-Ig

  Indirect ELISA and competitive ELISA results show that this antibody is specific to Pseudouridine. Indirect ELISA was performed by coating BSA conjugated Pseudouridine at 10ng/well followed by blocking with 1% BSA. Serial diluted primary antibody was added to the plates and incubated at 37℃. HRP-goat anti-mouse was used for detection. Competitive ELISA was performed similarly except that different concentration of Pseudouridine or its structure analogue compounds are mixed in primary antibody (fixed at 10 ng/mL).

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  Dot Blot experiment of RNA using 68578-1-Ig

  Total RNA was isolated from HeLa cell line and was dotted to NC membrane at different amount as indicated above the dots. The membrane was blocked with BSA and blotted with Pseudouridine antibody 68578-1-Ig at 1:2000 followed by incubation of HRP-goat anti-mouse secondary antibody. Signal was developed by ECL substrate. A parallel dot blot was performed using mouse IgG1 isotype control antibody 66360-1-Ig at the same dose.

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Tested Applications

• Positive ELISA detected in: Pseudouridine
• Positive Dot Blot detected in: RNA

Recommended dilution

• Enzyme-linked Immunosorbent Assay (ELISA): ELISA : 1:2000-1:20000
• DOT BLOT: DOT BLOT : 1:1000-1:4000

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

68578-1-Ig targets Pseudouridine in Dot Blot, ELISA applications and shows reactivity with chemical compound samples.

• Tested Reactivity: chemical compound
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: PTG
• Full Name: Pseudouridine
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Pseudouridine is an isomer of the nucleoside uridine in which the uracil is attached via a carbon-carbon instead of a nitrogen-carbon glycosidic bond. Pseudouridine is the most abundant RNA modification in cellular RNA (PMID: 29104216). After transcription and following synthesis, RNA can be modified with over 100 chemically distinct modifications. These can potentially regulate RNA expression post-transcriptionally, in addition to the four standard nucleotides and play a variety of roles in the cell including translation, localization and stabilization of RNA. Pseudouridine in rRNA and tRNA has been shown to fine-tune and stabilize the regional structure and help maintain their functions in mRNA decoding, ribosome assembly, processing and translation (PMID: 10902565, PMID: 23391857, PMID: 28045575). Pseudouridine in snRNA has been shown to enhance spliceosomal RNA-pre-mRNA interaction to facilitate splicing regulation (PMID 27268497).

Protocol for Dot Blot:
https://www.ptglab.com/protocol/68578-1-IgDotBlot.pdf