MABE419
Anti-m3G-cap, m7G-cap Antibody, clone H-20
clone H-20, from mouse
Synonym(s):
Anti-m3G/m7G-cap, Clone H-20 Anti-m3G-cap, m7G-cap Detection Antibody
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Recommended Products
biological source
mouse
Quality Level
antibody form
purified immunoglobulin
antibody product type
primary antibodies
clone
H-20, monoclonal
species reactivity (predicted by homology)
all
technique(s)
dot blot: suitable
immunocytochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable
isotype
IgG1κ
shipped in
wet ice
target post-translational modification
unmodified
General description
The nucleotide structure named the 2,2,7-trimethylguanosine(m3G)-containing cap structure of small nuclear ribonucleoprotein particles, or U snRNPs is an essential part of mRNA processing. Each snRNP particle consists of one (U1, U2 and U5) or two (U4/U6) snRNA molecules, a common set of eight core proteins (B, B9, D1, D2, D3, E, F and G, also denoted Sm proteins) that are bound to each of the 2,2,7- trimethylguanosine (m3G) cap-containing snRNAs U1, U2, U4 and U5, and several proteins associated specifically with the individual U snRNPs. Except for U6 snRNP, which does not leave the nucleus, the synthesis of these U snRNPs requires the bidirectional transport of the snRNA across the nuclear envelope. The snRNAs U1, U2, U4 and U5 are synthesized in the nucleus with a 7-monomethylguanosine (m7G) cap structure whereas the Sm proteins are stored in the cytoplasm and do not migrate into the nucleus in the absence of bound U snRNA. Instead, newly transcribed U snRNAs are transiently exported into the cytoplasm where the Sm proteins bind the snRNA’s Sm site, to form a ribonucleoprotein complex referred to as the Sm core. Stable association of all Sm proteins is essential for the hypermethylation of the m7G-cap to the m3G-cap structure. After this event and processing of the snRNAs, the mature snRNP particles are transported back to the nucleus in a receptor-and energy-dependent manner and the complete particle is formed. Monoclonal H-20 recognizes both m3G cap containing snRNPs as well as m7G capped structures and it should have a wide application, not only for studying the molecular biology and immunology of the U snRNPs from diverse organisms, but also for the characterization and isolation of m7G-capped transcripts.
Immunogen
Human m3G-cap and m7G-cap
Application
Anti-m3G-cap, m7G-cap Antibody, clone H-20 is a mouse monoclonal antibody, validated for use in Dot Blot, IP, western blotting & ICC.
Immunoprecipitation Analysis: A representative lot immunoprecipitated m3G-capped and m7G-capped Xenopus EF-1a, which was spiked into 4 µg of HeLa total RNA.
Western Blotting Analysis: A representative lot from an independent laboratory detected m3G-cap in m3G-capped snRNAs from HeLa nuclear extracts (Bochnig, P., et al. (1987). Eur J Biochem. 168(2):461-467.).
Immunoprecipitation Analysis: A representative lot from an independent laboratory immunoprecipitated m3G-cap from prefractionated HeLa S100 extracts (Huber, J., et al. (1998). EMBO J. 17(14):4114-4126.).
Immunocytochemistry Analysis: A representative lot from an independent laboratory detected m3G-cap in m3G-capped U1 snRNPs from digitonin permeabilized HeLa cells (Bochnig, P., et al. (1987). Eur J Biochem. 168(2):461-467.).
Western Blotting Analysis: A representative lot from an independent laboratory detected m3G-cap in m3G-capped snRNAs from HeLa nuclear extracts (Bochnig, P., et al. (1987). Eur J Biochem. 168(2):461-467.).
Immunoprecipitation Analysis: A representative lot from an independent laboratory immunoprecipitated m3G-cap from prefractionated HeLa S100 extracts (Huber, J., et al. (1998). EMBO J. 17(14):4114-4126.).
Immunocytochemistry Analysis: A representative lot from an independent laboratory detected m3G-cap in m3G-capped U1 snRNPs from digitonin permeabilized HeLa cells (Bochnig, P., et al. (1987). Eur J Biochem. 168(2):461-467.).
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
Chromatin Biology
Quality
Evaluated by Dot Blot in in vitro transcribed RNA without and with m7G-cap.
Dot Blot Analysis: 1 µg/mL of this antibody detected m7G-cap in in vitro transcribed RNA containing m7G-cap.
Dot Blot Analysis: 1 µg/mL of this antibody detected m7G-cap in in vitro transcribed RNA containing m7G-cap.
Physical form
Format: Purified
Protein G Purified
Purified mouse monoclonal IgG1κ in buffer containing PBS with 0.05% sodium azide.
Storage and Stability
Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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A monoclonal antibody against 2,2,7-trimethylguanosine that reacts with intact, class U, small nuclear ribonucleoproteins as well as with 7-methylguanosine-capped RNAs.
Bochnig, P, et al.
European Journal of Biochemistry, 168, 461-467 (1987)
Yasutoshi Akiyama et al.
RNA biology, 17(8), 1116-1124 (2020-03-03)
Recent transcriptome-wide studies have identified a diverse pool of transfer RNA (tRNA)-derived RNAs or tRNA-derived fragments (tRFs). Some of these RNAs have been demonstrated to be functional and involved in multiple biological processes ranging from the regulation of gene expression
Abhijit S Deshmukh et al.
Scientific reports, 6, 35288-35288 (2016-10-21)
Cyclin-dependent kinase 7 in conjunction with CyclinH and Mat1 activates cell cycle CDKs and is a part of the general transcription factor TFIIH. Role of Cdk7 is well characterized in model eukaryotes however its relevance in protozoan parasites has not
J Huber et al.
The EMBO journal, 17(14), 4114-4126 (1998-07-22)
The nuclear import of the spliceosomal snRNPs U1, U2, U4 and U5, is dependent on the presence of a complex nuclear localization signal (NLS). The latter is composed of the 5'-2,2,7-terminal trimethylguanosine (m3G) cap structure of the U snRNA and
Francesco Neri et al.
Nature, 543(7643), 72-77 (2017-02-23)
In mammals, DNA methylation occurs mainly at CpG dinucleotides. Methylation of the promoter suppresses gene expression, but the functional role of gene-body DNA methylation in highly expressed genes has yet to be clarified. Here we show that, in mouse embryonic
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