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RNA Methylation Essentials Antibody Kit

Host/Isotype

Rabbit/Mouse

Reactivity

Target Dependent

Applications

Target Dependent

Conjugate

Unconjugated

Type

Antibody Kit

Cat no : PK30016

Validation Data Gallery

Applications:
  • HEK-293 cells were lysised and immunoprecipitated with Protein A-m6A antibody and Protein A-mouse IgG3 control antibody respectively in the presence of RNAase inhibotor cocktail. The immunoprecipitated complex was washed diggested by RNAse A followed by western blot with YTHDF1(m6A reader) antibody 17479-1-AP (1:2000). (Lysate: 3.6mg per IP; IP: 15µg antibody and 50µL beads, 4 hours at 4℃; Diggestion: 50µg/mL * 80µL RNAse A for 1 hour at 37℃; Loading: 20% of elution; Input: 10µg.)

    HEK-293 cells were lysised and immunoprecipitated with Protein A-m6A antibody and Protein A-mouse IgG3 control antibody respectively in the presence of RNAase inhibotor cocktail. The immunoprecipitated complex was washed diggested by RNAse A followed by western blot with YTHDF1(m6A reader) antibody 17479-1-AP (1:2000). (Lysate: 3.6mg per IP; IP: 15µg antibody and 50µL beads, 4 hours at 4℃; Diggestion: 50µg/mL * 80µL RNAse A for 1 hour at 37℃; Loading: 20% of elution; Input: 10µg.)

  • HEK-293 cells were lysised and immunoprecipitated with Protein A-m6A antibody and Protein A-mouse IgG3 control antibody respectively in the presence of RNAase inhibotor cocktail. The immunoprecipitated complex was washed diggested by RNAse A followed by western blot with IGF2BP3 (m6A reader) antibody 66526-1-Ig (1:2000). (Lysate: 4.0 mg per IP; IP: 30µg antibody and 50µL beads, 4 hours at 4℃; Diggestion: 50µg/mL * 80µL RNAse A for 1 hour at 37℃; Loading: 20% of elution; Input: 10µg.)

    HEK-293 cells were lysised and immunoprecipitated with Protein A-m6A antibody and Protein A-mouse IgG3 control antibody respectively in the presence of RNAase inhibotor cocktail. The immunoprecipitated complex was washed diggested by RNAse A followed by western blot with IGF2BP3 (m6A reader) antibody 66526-1-Ig (1:2000). (Lysate: 4.0 mg per IP; IP: 30µg antibody and 50µL beads, 4 hours at 4℃; Diggestion: 50µg/mL * 80µL RNAse A for 1 hour at 37℃; Loading: 20% of elution; Input: 10µg.)

  • 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 1% BSA and blotted with m1A (1-Methyladenosine) antibody 68636-1-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 IgG2a isotype control Monoclonal antibody 66360-2-Ig at the same dose.

    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 1% BSA and blotted with m1A (1-Methyladenosine) antibody 68636-1-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 IgG2a isotype control Monoclonal antibody 66360-2-Ig at the same dose.

  • Total RNA was isolated from HEK-293 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 m6A antibody 68055-1-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 unrelated antibody with the same isotype (UCP2 antibody 66700-1-Ig) at the same dose.

    Total RNA was isolated from HEK-293 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 m6A antibody 68055-1-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 unrelated antibody with the same isotype (UCP2 antibody 66700-1-Ig) at the same dose.

  • Total DNA 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 m5C antibody 68301-1-Ig at 1:5000 followed by incubation of HRP-goat anti-mouse secondary antibody. Signal was developed by ECL substrate. A parallel dot blot was performed using Mouse IgG2b isotype control Monoclonal antibody 66360-3-Ig at the same dose.

    Total DNA 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 m5C antibody 68301-1-Ig at 1:5000 followed by incubation of HRP-goat anti-mouse secondary antibody. Signal was developed by ECL substrate. A parallel dot blot was performed using Mouse IgG2b isotype control Monoclonal antibody 66360-3-Ig at the same dose.

  • 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 m7G antibody 68302-1-Ig at 1:5000 followed by incubation of HRP-goat anti-mouse secondary antibody. Signal was developed by ECL substrate. A parallel dot blot was performed using Mouse IgG2b isotype control Monoclonal antibody 66360-3-Ig at the same dose.

    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 m7G antibody 68302-1-Ig at 1:5000 followed by incubation of HRP-goat anti-mouse secondary antibody. Signal was developed by ECL substrate. A parallel dot blot was performed using Mouse IgG2b isotype control Monoclonal antibody 66360-3-Ig at the same dose.

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 68302-1-Ig (m7G antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 68302-1-Ig (m7G antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 68302-1-Ig (m7G antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 68302-1-Ig (m7G antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 68302-1-Ig (m7G antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 68302-1-Ig (m7G antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 68055-1-Ig (m6A antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 68055-1-Ig (m6A antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 68055-1-Ig (m6A antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 68055-1-Ig (m6A antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 68301-1-Ig (m5C antibody) at dilution of 1:5000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 68301-1-Ig (m5C antibody) at dilution of 1:5000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 68301-1-Ig (m5C antibody) at dilution of 1:5000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 68301-1-Ig (m5C antibody) at dilution of 1:5000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

Product Information

The RNA Methylation Essentials Antibody Kit provides a cost-effective tool for studying the modifications m6A, m5C, m7G, and m1A. Perfect for researchers starting a new project, screening multiple prospective targets or those who simply require less volume. 

The RNA Methylation Essentials Antibody Kit contains antibodies against 4 key RNA modifications. 


AntigenCatalog No.Host, clonalityTested ReactivityApplicationsVolume
m6A68055-1-Ig         Mouse monoclonalH, M, R, Pg           WB, IP, IF, RIP, IHC, ELISA, Dot Blot20 uL
m5C68301-1-Ig         Mouse monoclonal            H, M, R       IHC, ELISA, Dot Blot20 uL
m7G68302-1-Ig         Mouse monoclonal             H, M        IHC, ELISA, Dot Blot           20 uL
m1A68636-1-Ig         Mouse monoclonal             H           ELISA, Dot Blot            20 uL


Also see our 'RNA Methylation Expanded Antibody Kit' on the following page  https://www.ptglab.com/products/RNA-Methylation-Expanded-Antibody-Kit-PK30017.htm

Storage

Store at -20°C. Stable for one year from the date of receipt. 

Background Information

RNA methylation is a type of RNA modification involving the addition of a methyl group to specific nucleotide bases. Methylation of RNA residues is modulated by the dynamic interplay between regulators called "writers" (methyltransferases), "readers" (binding proteins), and "erasers (demethylases). m6A, m5C, m7G, and m1A are common RNA modifications and have been shown to play critical roles in gene expression and various diseases, including cancer.  

Standard Protocols

Click  here  to view our standard protocols for various applications including WB, IP, IHC, IF, FC, and ELISA.

Cited in Article as

PK30016, RNA Methylation Essentials Antibody Kit, Proteintech, IL, USA