Recombinant Anti-METTL1 (phospho S27) antibody [EPR24280-9] (ab271062)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24280-9] to METTL1 (phospho S27)
- Suitable for: WB, Flow Cyt (Intra), ICC/IF, IP, Dot blot
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-METTL1 (phospho S27) antibody [EPR24280-9]
See all METTL1 primary antibodies -
Description
Rabbit monoclonal [EPR24280-9] to METTL1 (phospho S27) -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt (Intra), ICC/IF, IP, Dot blotmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293T, HeLa, RAW264.7 cell lysates ICC/IF: RAW 264.7, HeLa cells Flow Cyt: HeLa cells IP: HEK-293T cells
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24280-9 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab271062 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Predicted molecular weight: 31 kDa.
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Flow Cyt (Intra) |
1/500.
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ICC/IF |
1/50.
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IP |
1/30.
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Dot blot |
1/1000.
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Notes |
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WB
1/1000. Predicted molecular weight: 31 kDa. |
Flow Cyt (Intra)
1/500. |
ICC/IF
1/50. |
IP
1/30. |
Dot blot
1/1000. |
Target
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Function
Catalyzes the formation of N(7)-methylguanine at position 46 (m7G46) in tRNA. -
Tissue specificity
Ubiquitous. -
Pathway
tRNA modification; N(7)-methylguanine-tRNA biosynthesis. -
Sequence similarities
Belongs to the methyltransferase superfamily. TrmB family. -
Post-translational
modificationsPhosphorylation at Ser-27 inactivates its catalytic activity but does not affect the interaction with WDR4. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 4234 Human
- Entrez Gene: 17299 Mouse
- GenBank: BC000550 Human
- Omim: 604466 Human
- SwissProt: Q9UBP6 Human
- SwissProt: Q9Z120 Mouse
- Unigene: 42957 Human
- Unigene: 251593 Mouse
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Alternative names
- C12orf1 antibody
- D1075 like gene product antibody
- FLJ95748 antibody
see all
Images
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All lanes : Anti-METTL1 (phospho S27) antibody [EPR24280-9] (ab271062) at 1/1000 dilution
Lane 1 : Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : RAW264.7 treated with 100 nM Calycin A for 30 minutes, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution
Predicted band size: 31 kDa
Observed band size: 31 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 minutes
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All lanes : Anti-METTL1 (phospho S27) antibody [EPR24280-9] (ab271062) at 1/1000 dilution
Lane 1 : Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa treated with 100 nM Calycin A for 30 minutes, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution
Predicted band size: 31 kDa
Observed band size: 31 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 minutes
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All lanes : Anti-METTL1 (phospho S27) antibody [EPR24280-9] (ab271062) at 1/1000 dilution
Lane 1 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate (Untreated membrane)
Lane 2 : HEK-293T treated with 100 nM Calycin A for 30 minutes, whole cell lysate (Untreated membrane)
Lane 3 : HEK-293T treated with /ml PDGF-AA for 5 minutes then treated with 100 nM Calyculin A for 30 minutes, whole cell lysate (Untreated membrane)
Lane 4 : HEK-293T transfected with an AKT1 expression vector, and treated with 100 ng/ml PDGF-AA for 5 minutes then treated with 100 nM Calyculin A for 30 minutes, whole cell lysate (Untreated membrane)
Lane 5 : HEK-293T whole cell lysate (phosphatase treated membrane)
Lane 6 : HEK-293T treated with 100 nM Calycin A for 30 minutes, whole cell lysate (phosphatase treated membrane)
Lane 7 : HEK-293T treated with /ml PDGF-AA for 5 minutes then treated with 100 nM Calyculin A for 30 minutes, whole cell lysate (phosphatase treated membrane)
Lane 8 : HEK-293T transfected with an AKT1 expression vector, and treated with /ml PDGF-AA for 5 minutes then treated with 100 nM Calyculin A for 30 minutes, whole cell lysate (phosphatase treated membrane)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 31 kDa
Observed band size: 31 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBSTAKT1 in lane 4 and lane 8 act as a kinase to active METTL1 (phospho S27).
Exposure time: 26 seconds
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Dot blot analysis of METTL1 (phospho S27) using ab271062 at 1:1000 (0.547 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1:100,000 dilution.
Exposure time: 3 minutes
Blocking and diluting buffer and concentration: 5% NFDM/TBST
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METTL1 (phospho S27) was immunoprecipitated from 0.35 mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100 nM Calyculin A for 30 minutes, whole cell lysate with ab271062 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271062 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100 nM Calyculin A for 30 minutes, whole cell lysate 10 ug
Lane 2: ab271062 IP in RAW264.7 treated with 100 nM Calyculin A for 30 minutes, whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab271062 in RAW264.7 treated with 100 nM Calyculin A for 30 minutes, whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 50 seconds
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METTL1 (phospho S27) was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney epithelial cell) transfected with an AKT1 expression vector, and treated with 100ng/ml PDGF-AA for 5 minutes then treated with 100 nM Calyculin A for 30 minutes, whole cell lysate with ab271062 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271062 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HEK-293T (human embryonic kidney epithelial cell) transfected with an AKT1 expression vector, and treated with 100ng/ml PDGF-AA for 5 minutes then treated with 100 nM Calyculin A for 30 minutes, whole cell lysate 10 ug
Lane 2: ab271062 IP in HEK-293T transfected with an AKT1 expression vector, and treated with 100ng/ml PDGF-AA for 5 minutes then treated with 100 nM Calyculin A for 30 minutes, whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab271062 in HEK-293T transfected with an AKT1 expression vector, and treated with 100ng/ml PDGF-AA for 5 minutes then treated with 100 nM Calyculin A for 30 minutes, whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3.25 seconds
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100nM Calyculin A for 5min (Red) / Untreated control (Green) cells labelling METTL1 (phospho S27) with ab271062 at 1/500 dilution (0.1ug)/ Red and Green compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) treated with 100nM Calyculin A for 5min (Red) / Untreated control (Green) cells labelling METTL1 (phospho S27) with with ab271062 at 1/500 dilution (0.1ug)/ Red and Green compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling METTL1 (phospho S27) with ab271062 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing increased cytoplasmic and nuclear staining in HeLa cells treated with Calyculin A (100 nM) for 5 min. is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling METTL1 (phospho S27) with ab271062 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing increased cytoplasmic and nuclear staining in RAW 264.7 cells treated with Calyculin A (100 nM) for 5 min. is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab271062 has not yet been referenced specifically in any publications.