Recombinant Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free (ab277533)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24020-38] to NPTX2 - BSA and Azide free
- Suitable for: ICC/IF, IHC-P, WB, IHC-Fr, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-NPTX2 antibody [EPR24020-38] - BSA and Azide free
See all NPTX2 primary antibodies -
Description
Rabbit monoclonal [EPR24020-38] to NPTX2 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IHC-P, WB, IHC-Fr, IP, Flow Cyt (Intra)more 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: Human brain, pancreas and testis tissue lysates; SH-SY5Y whole cell lysate; Mouse brain, kidney and testis tissue lysates; Rat brain and testis tissue lysates. IHC-P: Human pancreas and cerebrum tissue; Mouse cerebrum tissue; Rat cerebrum tissue. ICC/IF: Beta-TC-6 cells. Flow Cyt (intra): Beta-TC-6 cells. IP: Human brain tissue lysate; Mouse brain tissue lysate. IHC-Fr: Frozen human cerebral cortex, pancreas, and heart tissue sections.
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General notes
ab277533 is the carrier-free version of ab277523.
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
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24020-38 -
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 ab277533 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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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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WB |
Use at an assay dependent concentration. Detects a band of approximately 50-60 kDa (predicted molecular weight: 47 kDa).
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IHC-Fr |
Use at an assay dependent concentration.
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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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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. |
WB
Use at an assay dependent concentration. Detects a band of approximately 50-60 kDa (predicted molecular weight: 47 kDa). |
IHC-Fr
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
Target
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Function
Likely to play role in the modification of cellular properties that underlie long-term plasticity. Binds to agar matrix in a calcium-dependent manner. -
Tissue specificity
Brain, pancreas, liver, heart and skeletal muscle. Highest levels are seen in the testis. -
Sequence similarities
Contains 1 pentaxin domain. -
Cellular localization
Secreted. - Information by UniProt
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Database links
- Entrez Gene: 4885 Human
- Entrez Gene: 53324 Mouse
- Entrez Gene: 288475 Rat
- Omim: 600750 Human
- SwissProt: P47972 Human
- SwissProt: O70340 Mouse
- SwissProt: P97738 Rat
- Unigene: 3281 Human
see all -
Alternative names
- apexin antibody
- NARP antibody
- neuronal activity-regulated pentaxin antibody
see all
Images
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This data was developed using ab277523, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling NPTX2 with ab277523 at 1/2000 (0.229 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on mouse cerebrum (PMID:31772567). The section was incubated with ab277523 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 ab277523, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling NPTX2 with ab277523 at 1/2000 (0.229 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Positive staining on human cerebrum. The section was incubated with ab277523 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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This data was developed using ab277523, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 100% Methanol permeabilized Beta-TC-6 cells labelling NPTX2 with ab277523 at 1/50 (9.16 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor®488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Beta-TC-6 cells. 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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All lanes : Anti-NPTX2 antibody [EPR24020-38] (ab277523) at 1/1000 dilution
Lane 1 : Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Mouse testis tissue lysate
Lane 4 : Mouse kidney tissue lysate
Lane 5 : Mouse lung tissue lysate
Lane 6 : Rat brain tissue lysate
Lane 7 : Rat testis tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 47 kDa
Observed band size: 50-60 kDa why is the actual band size different from the predicted?This data was developed using ab277523, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
NPTX2 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID: 8786423, 28440221).
Exposure time: 15 seconds.
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This data was developed using ab277523, the same antibody clone in a different buffer formulation.
NPTX2 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab277523 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab277523 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Mouse brain tissue lysate 10 ug
Lane 2: ab277523 IP in Mouse brain tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab277523 in mouse brain tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
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This data was developed using ab277523, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Beta-TC-6 (Mouse pancreas insulinoma beta cell) cells labelling NPTX2 with ab277523 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730)isotype control (Black) 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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IHC image of NPTX2 staining in a section of frozen normal human cerebral cortex performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab277523, 0.5 ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.This data was developed using ab277523, the same antibody clone in a different buffer formulation.
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IHC image of NPTX2 staining in a section of frozen normal human pancreas performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab277523, 0.5 ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.This data was developed using ab277523, the same antibody clone in a different buffer formulation.
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Negative tissue image: IHC image of NPTX2 staining in a section of frozen normal human heart performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab277523, 0.5 ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.This data was developed using ab277523, the same antibody clone in a different buffer formulation.
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This data was developed using ab277523, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling NPTX2 with ab277523 at 1/2000 (0.229 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on islet of human pancreas. The section was incubated with ab277523 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-NPTX2 antibody [EPR24020-38] (ab277523) at 1/1000 dilution
Lane 1 : Human brain tissue lysate
Lane 2 : Human pancreas tissue lysate
Lane 3 : Human testis tissue lysate
Lane 4 : Human kidney tissue lysate
Lane 5 : Human lung tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution
Predicted band size: 47 kDa
Observed band size: 50-60 kDa why is the actual band size different from the predicted?This data was developed using ab277523, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
NPTX2 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID: 8786423, 28440221).
Negative: human kidney, human lung (PMID: 8530029, 8786423).
Exposure times: Lane 1: 15 secondsLane 2-5: 3 minutes.
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This data was developed using ab277523, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling NPTX2 with ab277523 at 1/2000 (0.229 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on rat cerebrum. The section was incubated with ab277523 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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Anti-NPTX2 antibody [EPR24020-38] (ab277523) at 1/1000 dilution + SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 47 kDa
Observed band size: 50-60 kDa why is the actual band size different from the predicted?This data was developed using ab277523, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
NPTX2 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID: 8786423, 28440221).
Exposure time: 15 seconds.
-
This data was developed using ab277523, the same antibody clone in a different buffer formulation.
NPTX2 was immunoprecipitated from 0.35 mg Human brain tissue lysate with ab277523 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab277523 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Human brain tissue lysate 10 ug
Lane 2: ab277523 IP in Human brain tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab277523 in human brain tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 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)
ab277533 has not yet been referenced specifically in any publications.