N4-acetylcytidine Monoclonal antibody

N4-acetylcytidine Monoclonal Antibody for Dot Blot, ELISA

Host / Isotype

Mouse / IgG1

Reactivity

chemical compound

Applications

Dot Blot, ELISA

Conjugate

Unconjugated

CloneNo.

3C2C2

Cat no : 68498-1-Ig

Synonyms

C11H15N3O6, Ac-rC, Acetylcytidine, ACETYL CYTOSINE, Ac4C

---

Validation Data Gallery

• 

  ELISA experiment of N4-acetylcytidine using 68498-1-Ig

  Indirect ELISA and competitive ELISA results show that this antibody is specific to ac4C (N4-acetylcytidine). Indirect ELISA (blue curve, refer to top X-right Y axis) was performed by coating BSA conjugated ac4C at 50ng/well followed by blocking with 5% non fat milk. 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 Ac4C or its structure analogue compounds was mixed in 20ng/mL primary antibody (red curve, refer to bottom X-left Y axis).

• 

  Dot Blot experiment of HeLa using 68498-1-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 5% milk and blotted with N4-acetylcytidine antibody 68498-1-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 Monoclonal antibody 66360-1-Ig at the same dose.

---

Tested Applications

• Positive ELISA detected in: N4-acetylcytidine
• Positive Dot Blot detected in: RNA isolate HeLa cells

Recommended dilution

• Enzyme-linked Immunosorbent Assay (ELISA): ELISA :
• 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

68498-1-Ig targets N4-acetylcytidine 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: Compound
• Full Name: N4-acetylcytidine
• 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

N4-Acetylcytidine, CasNo. 3768-18-1, is a modified nucleoside and endogenous urinary nucleoside product of the degradation of tRNA, 18s rRNA and mRNA. N4-Acetylcytidine is a biological marker for various cancers with elevated concentrations present in urine. N4-Acetylcytidine is also a partially protected cytidine and therefore can be used as a synthetic building block to prepare further derivatized nucleosides such as 2',3'-dideoxycytidine. NAT10 catalyzes the formation of N4-acetylcytidine (ac4C) modification on mRNAs, 18S rRNA and tRNAs.

Protocol for Dot Blot:
https://www.ptglab.com/protocol/68498-1-IgDotBlot.pdf