m7G Monoclonal antibody

m7G Monoclonal Antibody for IHC, Dot Blot, ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

chemical compound and More (1)

Applications

IHC, Dot Blot, ELISA

Conjugate

Unconjugated

Publications

1

CloneNo.

2C3E10

Cat no : 68302-1-Ig

Synonyms

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Validation Data Gallery

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  IHC staining of human colon cancer using 68302-1-Ig

  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).

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  IHC staining of human pancreas cancer using 68302-1-Ig

  Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 68302-1-Ig (m7G antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver cancer using 68302-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 68302-1-Ig (m7G antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse testis using 68302-1-Ig

  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).

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  IHC staining of mouse testis using 68302-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 68302-1-Ig (m7G antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  ELISA experiment of compound using 68302-1-Ig

  Indirect ELISA and competitive ELISA results show that this antibody is specific to m7G. Indirect ELISA (blue curve, refer to top X-right Y axis) was performed by coating BSA conjugated m7G 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 m7G or its structure analogue compound Guanosine was mixed in 20ng/mL primary antibody (red curve, refer to bottom X-left Y axis).

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  Dot Blot experiment of HeLa using 68302-1-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 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.

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Tested Applications

• Positive IHC detected in: mouse testis tissue, human colon cancer tissue, human liver cancer tissue, human pancreas cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive ELISA detected in: compound
• Positive Dot Blot detected in: HeLa cells

Recommended dilution

• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Enzyme-linked Immunosorbent Assay (ELISA): ELISA : 1:10-1:100
• DOT BLOT: DOT BLOT : 1:2500-1:10000

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

68302-1-Ig targets m7G in IHC, Dot Blot, ELISA applications and shows reactivity with chemical compound samples.

• Tested Reactivity: chemical compound
• Cited Reactivity: mouse
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: compound
• Full Name: m7G
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A 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

7-Methylguanosine (m7G) is a modified purine nucleoside. It is a methylated version of guanosine and when found in human urine, it may be a biomarker of some types of cancer. In the RNAs, 7-methylguanosine have been used to study and examine the reaction evolving methylguanosine. It also plays a role in mRNA as a blocking group at its 5´-end. The m7G modification actively participates in biological and pathological functions by affecting the metabolism of various RNA molecules, including messenger RNA, ribosomal RNA, microRNA, and transfer RNA. Increasing evidence indicates a critical role for m7G in human disease development, especially cancer, and aberrant m7G levels are closely associated with tumorigenesis and progression via regulation of the expression of multiple oncogenes and tumor suppressor genes.

Protocol for Dot Blot:
https://www.ptglab.com/protocol/68302-1-IgDotBlot.pdf

Publications

Species	Application	Title
mouse		Eur J Med Res; A comprehensive analysis of m6A/m7G/m5C/m1A-related gene expression and immune infiltration in liver ischemia-reperfusion injury by integrating bioinformatics and machine learning algorithms; Authors - Zhanzhi Meng