Recombinant Anti-NOP2 antibody [EPR24888-92] - BSA and Azide free (ab282822)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24888-92] to NOP2 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-NOP2 antibody [EPR24888-92] - BSA and Azide free
See all NOP2 primary antibodies -
Description
Rabbit monoclonal [EPR24888-92] to NOP2 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, 293T, NIH/3T3 and PC-12 whole cell lysate. IHC-P: Human cerebrum, Human lung adenocarcinoma, Mouse cerebrum and Rat cerebrum tissue. ICC/IF: HCT 116 and Neuro-2a cells. Flow Cyt(Intra): HCT 116 cells. IP: 293T cells.
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General notes
ab282822 is the carrier-free version of ab271075.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. -
Storage buffer
pH: 7.2
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24888-92 -
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 ab282822 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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 89 kDa.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 89 kDa. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
Target
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Relevance
NOP2 belongs to the methyltransferase superfamily of the RsmB/NOP family. NOP2 may play a role in the regulation of the cell cycle and the increased nucleolar activity that is associated with the cell proliferation. May act as ribosomal RNA methyltransferase. -
Cellular localization
Nuclear; nucleolus -
Database links
- Entrez Gene: 4839 Human
- Entrez Gene: 110109 Mouse
- Entrez Gene: 314969 Rat
- Omim: 164031 Human
- SwissProt: P46087 Human
- SwissProt: Q922K7 Mouse
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Alternative names
- NSUN2 antibody
- MISU antibody
- NOL1 antibody
see all
Images
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All lanes : Anti-NOP2 antibody [EPR24888-92] (ab271075) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : 293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 89 kDa
Observed band size: 110,90 kDa why is the actual band size different from the predicted?This data was developed using ab271075, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Lysates were made freshly and used in WB test immediately to minimize protein degradation.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:25043274).
The multiple reactive bands would represent different NOP2 isoforms.
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This data was developed using ab271075, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labelling NOP2 with ab271075 at 1/200 dilution (2.835 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nucleolar staining on human cerebrum (PMID: 25481415). The section was incubated with ab271075 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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This data was developed using ab271075, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma tissue labelling NOP2 with ab271075 at 1/200 dilution (2.835 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on human lung adenocarcinoma (PMID: 3422591). The section was incubated with ab271075 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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This data was developed using ab271075, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labelling NOP2 with ab271075 at 1/500 dilution (1.134 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nucleolar staining on mouse cerebrum. The section was incubated with ab271075 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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This data was developed using ab271075, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labelling NOP2 with ab271075 at 1/500 dilution (1.134 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nucleolar staining on rat cerebrum. The section was incubated with ab271075 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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This data was developed using ab271075, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HCT 116 (human colorectal carcinoma cell line) cells labelling NOP2 with ab271075 at 1/500 dilution (1.134 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml / Green). Confocal image showing nucleolar staining in HCT 116 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/ml / Red). 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 (2 µg/ml) dilution.
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This data was developed using ab271075, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (Mouse neuroblastoma cell line) cells labelling NOP2 with ab271075 at 1/500 dilution (1.134 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml / Green). Confocal image showing nucleolar staining in Neuro-2a cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/ml / Red). 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 (2 µg/ml).
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This data was developed using ab271075, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cells labelling NOP2 with ab271075 at 1/50 dilution (1µg / Red) 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, ab150077) at 1/2000 dilution was used as the secondary antibody.
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This data was developed using ab271075, the same antibody clone in a different buffer formulation.
NOP2 was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) whole cell lysate with ab271075 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271075 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: 293T (human embryonic kidney epithelial cell) whole cell lysate 10 µg
Lane 2: ab271075 IP in 293T whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab271075 in 293T whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 24 seconds
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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Datasheet download
Certificate of Compliance
References (0)
ab282822 has not yet been referenced specifically in any publications.