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m6A Recombinant Rabbit Monoclonal Antibody [1G1] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750493

m6A Recombinant Rabbit Monoclonal Antibody [1G1] - BSA and Azide free

Application

  • Dot Blot

  • ELISA

Reactivity

  • Species independent

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

m6A Recombinant Rabbit Monoclonal Antibody [1G1] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

m6A Small Molecule conjugated to OVA.

Species Reactivity

Species independent

Validated Applications

Dot Blot, ELISA

Conjugation

unconjugated

Clone Number

1G1

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • Dot Blot

  • 1:500-1:2,000

  • ELISA

  • 1:1,000-1:2,000

Target

Function

N6-methyladenosine (m6A) is the most prevalent and abundant post-transcriptional RNA modification on eukaryote mRNA, and its biological functions are mediated by special binding proteins (i.e., methyltransferases, demethylases, and effectors) that recognize this modification. The presence of m6A on transcripts contributes to diverse fundamental cellular functions, such as pre-mRNA splicing, nuclear transport, stability, translation, and microRNA biogenesis, implying an association with numerous human diseases. The linkages between m6A and numerous cancer types have been indicated in reports that include stomach cancer, prostate cancer, breast cancer, pancreatic cancer, kidney cancer, mesothelioma, sarcoma, and leukaemia. The impacts of m6A on cancer cell proliferation might be much more profound with more data emerging. The depletion of METTL3 is known to cause apoptosis of cancer cells and reduce invasiveness of cancer cells, while the activation of ALKBH5 by hypoxia was shown to cause cancer stem cell enrichment. m6A has also been indicated in the regulation of energy homeostasis and obesity, as FTO is a key regulatory gene for energy metabolism and obesity. SNPs of FTO have been shown to associate with body mass index in human populations and occurrence of obesity and diabetes. The influence of FTO on pre-adipocyte differentiation has been suggested. The connection between m6A and neuronal disorders has also been studied. For instance, neurodegenerative diseases may be affected by m6A as the cognate dopamine signalling was shown to be dependent on FTO and correct m6A methylation on key signalling transcripts. The mutations in HNRNPA2B1, a potential reader of m6A, have been known to cause neurodegeneration. The IGF2BP1–3, a novel class of m6A reader, has oncogenic functions. IGF2BP1–3 knockdown or knockout decreased MYC protein expression, cell proliferation and colony formation in human cancer cell lines. The ZC3H13, a member of the m6A methyltransferase complex, markedly inhibited colorectal cancer cells growth when knocked down.

Background References

1. Sun T. et. al. The role of m6A RNA methylation in cancer. Biomed Pharmacother. 2019 Apr

2. Qin Y. et. al. Role of m6A RNA methylation in cardiovascular disease (Review). Int J Mol Med. 2020 Dec

Synonyms

m6A antibody

N6-methyladenosine antibody

Images

  • Dot blot analysis of N6-methyladenosine (m6A) using HA750493.<br /><br />Loading: total RNA extracted from HeLa<br />Blocking Buffer: 5% NFDM/TBST;<br />Incubation time and temperature: 1 hours at room temperature;<br />Primary Antibody: HA750493<br />Dilution: 2 ug/ml, 1 ug/ml, 0.5 ug/ml, 0.25 ug/ml, 0.125 ug/ml, 0.0625 ug/ml<br />Dilution Buffer: Primary Antibody Dilution Buffer form Beyotime;<br />Incubation Time and Temperature: Overnight at 4 ℃ ;<br />Secondary Antibody: Goat anti-Rabbit IgG-HRP antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>);<br />Dilution: 1:300,000 <br />Dilution Buffer: 5% NFDM/TBST;<br />Incubation Time and Temperature: 30 minutes at room temperature;<br />Exposure time: 60/120 seconds.

    Dot blot analysis of N6-methyladenosine (m6A) using HA750493.

    Loading: total RNA extracted from HeLa
    Blocking Buffer: 5% NFDM/TBST;
    Incubation time and temperature: 1 hours at room temperature;
    Primary Antibody: HA750493
    Dilution: 2 ug/ml, 1 ug/ml, 0.5 ug/ml, 0.25 ug/ml, 0.125 ug/ml, 0.0625 ug/ml
    Dilution Buffer: Primary Antibody Dilution Buffer form Beyotime;
    Incubation Time and Temperature: Overnight at 4 ℃ ;
    Secondary Antibody: Goat anti-Rabbit IgG-HRP antibody (HA1001);
    Dilution: 1:300,000
    Dilution Buffer: 5% NFDM/TBST;
    Incubation Time and Temperature: 30 minutes at room temperature;
    Exposure time: 60/120 seconds.

  • Competitive ELISA analysis of m6A was performed by coating wells of a 96-well plate with 50 µl per well of m6A -BSA diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 1%BSA blocking buffer, and incubated with 100 µl per well of m6A monoclonal antibody at concentration of 1 µg/mL with serial diluted m6A and its analogs starting from a concentration of 10,000ng/ml to 78ng/ml for 1 hours at room temperature. The plate was washed and incubated with 50 µl per well of an HRP-conjugated goat anti-Rabbit IgG secondary antibody at a dilution of 1:15,000 for one hour at room temperature. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

    Competitive ELISA analysis of m6A was performed by coating wells of a 96-well plate with 50 µl per well of m6A -BSA diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 1%BSA blocking buffer, and incubated with 100 µl per well of m6A monoclonal antibody at concentration of 1 µg/mL with serial diluted m6A and its analogs starting from a concentration of 10,000ng/ml to 78ng/ml for 1 hours at room temperature. The plate was washed and incubated with 50 µl per well of an HRP-conjugated goat anti-Rabbit IgG secondary antibody at a dilution of 1:15,000 for one hour at room temperature. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

  • Competitive ELISA analysis of m6A was performed by coating wells of a 96-well plate with 50 µl per well of m6A -BSA diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 1%BSA blocking buffer, and incubated with 100 µl per well of m6A monoclonal antibody at concentration of 1 µg/mL with m6A and its analogs at a concentration of 10ug/ml for 1 hours at room temperature. The plate was washed and incubated with 50 µl per well of an HRP-conjugated goat anti-Rabbit IgG secondary antibody at a dilution of 1:15,000 for one hour at room temperature. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

    Competitive ELISA analysis of m6A was performed by coating wells of a 96-well plate with 50 µl per well of m6A -BSA diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 1%BSA blocking buffer, and incubated with 100 µl per well of m6A monoclonal antibody at concentration of 1 µg/mL with m6A and its analogs at a concentration of 10ug/ml for 1 hours at room temperature. The plate was washed and incubated with 50 µl per well of an HRP-conjugated goat anti-Rabbit IgG secondary antibody at a dilution of 1:15,000 for one hour at room temperature. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"