Recombinant Anti-NOL3 antibody [EPR25182-11] - BSA and Azide free (ab288304)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR25182-11] to NOL3 - BSA and Azide free
- Suitable for: WB, ICC/IF, IHC-P, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-NOL3 antibody [EPR25182-11] - BSA and Azide free
See all NOL3 primary antibodies -
Description
Rabbit monoclonal [EPR25182-11] to NOL3 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, IHC-P, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MCF7 whole cell lysate; Human heart and skeletal muscle tissue lysates; Mouse heart and skeletal muscle tissue lysates; Rat heart and skeletal muscle tissue lysates. IHC-P: Human skeletal muscle tissue; Mouse cardiac muscle tissue; Rat cardiac muscle tissue. ICC/IF: MCF7 cells. Flow cyt: MCF7 cells. IP: MCF7 and Neuro-2a whole cell lysates.
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General notes
ab288304 is the carrier-free version of ab288295.
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
EPR25182-11 -
Isotype
IgG -
Research areas
Associated products
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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 ab288304 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 |
Use at an assay dependent concentration. Detects a band of approximately 27 kDa (predicted molecular weight: 24 kDa).
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ICC/IF |
Use at an assay dependent concentration.
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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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. Detects a band of approximately 27 kDa (predicted molecular weight: 24 kDa). |
ICC/IF
Use at an assay dependent concentration. |
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. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
Target
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Function
Isoform 1 may be involved in RNA splicing.
Isoform 2 may inhibit apoptosis. -
Tissue specificity
Highly expressed in heart and skeletal muscle. Detected at low levels in placenta, liver, kidney and pancreas. -
Sequence similarities
Contains 1 CARD domain. -
Cellular localization
Cytoplasm and Nucleus > nucleolus. - Information by UniProt
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Database links
- Entrez Gene: 8996 Human
- Entrez Gene: 78688 Mouse
- Entrez Gene: 85383 Rat
- Omim: 605235 Human
- SwissProt: O60936 Human
- SwissProt: Q9D1X0 Mouse
- SwissProt: Q62881 Rat
- Unigene: 513667 Human
see all -
Alternative names
- Apoptosis repressor with CARD antibody
- ARC antibody
- Muscle enriched cytoplasmic protein antibody
see all
Images
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This data was developed using ab288295, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labelling NOL3 with ab288295 at 1/5000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond®, Polymer Refine Detection). Positive staining on rat cardiac muscle. The section was incubated with ab288295 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).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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All lanes : Anti-NOL3 antibody [EPR25182-11] (ab288295) at 1/1000 dilution
Lane 1 : Mouse heart tissue lysate
Lane 2 : Mouse liver tissue lysate
Lane 3 : Mouse skeletal muscle tissue lysate
Lane 4 : Rat heart tissue lysate
Lane 5 : Rat skeletal muscle tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 24 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?This data was developed using ab288295, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer: 5% NFDM/TBST.
The observed MW/expression profile is consistent with what has been described in the literature (PMID: 14645204, 20026055, 10590251).
Low expression: Mouse liver (PMID: 14645204)
Exposure time: 3 minutes
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This data was developed using ab288295, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Daudi (Human Burkitt's lymphoma lymphoblast, Left) / MCF7 (Human breast adenocarcinoma epithelial cell, Right) cells labelling NOL3 with ab288295 at 1/5000 dilution (0.01ug)(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, ab150081) at 1/2000 dilution was used as the secondary antibody.
Negative control: Daudi (PMID: HPA database).
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This data was developed using ab288295, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a cells labelling NOL3 with ab288295 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in Neuro-2a cell line. 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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All lanes : Anti-NOL3 antibody [EPR25182-11] (ab288295) at 1/1000 dilution
Lane 1 : Human heart tissue lysate
Lane 2 : Human skeletal muscle tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 24 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?This data was developed using ab288295, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The observed MW/expression profile is consistent with what has been described in the literature (PMID: 15848180).
Exposure time: 15 seconds
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All lanes : Anti-NOL3 antibody [EPR25182-11] (ab288295) at 1/1000 dilution
Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : Daudi (human Burkitts lymphoma lymphoblast) 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: 24 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?This data was developed using ab288295, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The observed MW/expression profile is consistent with what has been described in the literature (PMID: 17142452).
Negative control: Daudi (PMID: HPA database)
Exposure time: 3 minutes
-
This data was developed using ab288295, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labelling NOL3 with ab288295 at 1/5000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond®, Polymer Refine Detection). Positive staining on mouse cardiac muscle. The section was incubated with ab288295 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).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
This data was developed using ab288295, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling NOL3 with ab288295 at 1/5000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Cytoplasmic staining on human skeletal muscle. The section was incubated with ab288295 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).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
This data was developed using ab288295, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling NOL3 with ab288295 at 1/5000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond®, Polymer Refine Detection) was used. Negative control: No staining on mouse liver. The section was incubated with ab288295 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
-
This data was developed using ab288295, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MCF7 cells labelling NOL3 with ab288295 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in MCF7 cell line. Negative control: Daudi. 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.
-
This data was developed using ab288295, the same antibody clone in a different buffer formulation.
NOL3 was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with ab288295 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288295 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 ug
Lane 2: ab288295 IP in MCF7 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab288295 in MCF7 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds
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This data was developed using ab288295, the same antibody clone in a different buffer formulation.
NOL3 was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab288295 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288295 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate 10 ug
Lane 2: ab288295 IP in Neuro-2a whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab288295 in Neuro-2a whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
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)
ab288304 has not yet been referenced specifically in any publications.