Anti-Nestin antibody [Rat-401] - Neural Stem Cell Marker (ab6142)
Key features and details
- Mouse monoclonal [Rat-401] to Nestin - Neural Stem Cell Marker
- Suitable for: IHC-P, WB
- Reacts with: Mouse, Rat
- Isotype: IgG1
Related conjugates and formulations
Overview
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Product name
Anti-Nestin antibody [Rat-401] - Neural Stem Cell Marker
See all Nestin primary antibodies -
Description
Mouse monoclonal [Rat-401] to Nestin - Neural Stem Cell Marker -
Host species
Mouse -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat
Does not react with: Sheep, Cat, Monkey -
Immunogen
Tissue, cells or virus corresponding to Rat Nestin. Homogenized spinal cord tissue from embryonic day 15 (E15) rats.
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Positive control
- WB: Mouse brain and rat brain whole tissue lysates. IHC-P: Rat Brain 6 weeks (cerebellum sagittal, coronal rest) tissue sections.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
Rat-401 -
Isotype
IgG1 -
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 ab6142 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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IHC-P | (8) |
Use a concentration of 0.05 - 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Tissue fixed with 4% paraformaldehyde at pH 7.4 for light microscopy. |
WB | (3) |
Use a concentration of 1 µg/ml. Predicted molecular weight: 200 kDa.
Block with milk or BSA but do not dilute primary antibody in buffer containing milk. |
Notes |
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IHC-P
Use a concentration of 0.05 - 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Tissue fixed with 4% paraformaldehyde at pH 7.4 for light microscopy. |
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 200 kDa. Block with milk or BSA but do not dilute primary antibody in buffer containing milk. |
Target
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Function
Required for brain and eye development. Promotes the disassembly of phosphorylated vimentin intermediate filaments (IF) during mitosis and may play a role in the trafficking and distribution of IF proteins and other cellular factors to daughter cells during progenitor cell division. Required for survival, renewal and mitogen-stimulated proliferation of neural progenitor cells. -
Tissue specificity
CNS stem cells. -
Sequence similarities
Belongs to the intermediate filament family. -
Developmental stage
Upon terminal neural differentiation, nestin is down-regulated and replaced by neurofilaments. -
Post-translational
modificationsConstitutively phosphorylated. This increases during mitosis when the cytoplasmic intermediate filament network is reorganized. - Information by UniProt
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Database links
- Entrez Gene: 18008 Mouse
- SwissProt: Q6P5H2 Mouse
- SwissProt: P21263 Rat
- Unigene: 331129 Mouse
- Unigene: 9701 Rat
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Alternative names
- ESTM 46 antibody
- FLJ 21841 antibody
- FLJ21841 antibody
see all
Images
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IHC image of Nestin staining in a section of formalin-fixed paraffin-embedded normal rat brain performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab6142, 0.05ug/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 hematoxylin and mounted with DPX.
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.
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All lanes : Anti-Nestin antibody [Rat-401] - Neural Stem Cell Marker (ab6142) at 1 µg/ml
Lane 1 : Mouse E12 brain tissue lysate
Lane 2 : Mouse E14 brain tissue lysate
Lane 3 : Mouse E16 brain tissue lysate
Lane 4 : Mouse E18 brain tissue lysate
Lane 5 : Rat E12 brain tissue lysate
Lane 6 : Rat E14 brain tissue lysate
Lane 7 : Rat E16 brain tissue lysate
Lane 8 : Rat E18 brain tissue lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 200 kDaThis blot was produced using a 3-8% Tris-Acetate gel under the Tris-Acetate buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab6142 overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution for 1 hour at room temperature before imaging.
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REST expression in N-hREST mouse brains correlates with stemness in embryonic neural stem cells. Immunofluorescence analysis of E18.5 N-hREST and LSL-hRESTcontrol littermate mouse brains with antibodies against REST (using an antibody that preferentially recognizes hREST over mouse REST) and Nestin (using ab6142)
Mice were anesthetized and perfused with phosphate-buffered saline followed by 4% paraformaldehyde (PFA). Brain tissues were then dissected and fixed in 4% PFA overnight at 4 °C. Fixed brain tissues were processed for paraffin embedding and then cut into 5-μm sections.
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ab6142 staining adult mouse brain tissue section by Immunohistochemistry (Formalin/PFA-fixed, paraffin embedded sections). Tissue underwent fixation in paraformaldehyde, heat mediated antigen retrieval in Sodium Citrate, permeabilization in 1% Triton buffer and blocking in 10% serum for 1 hour at 25°C. The primary antibody, diluted 1/200 (PBS, 2% Donkey serum, 0.2% Triton) for 16 hours at 4°C. An Alexa Fluor® 488 conjugated donkey polyclonal to mouse Ig, diluted 1/500 was used as the secondary.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (209)
ab6142 has been referenced in 209 publications.
- Romero DM et al. Novel role of the synaptic scaffold protein Dlgap4 in ventricular surface integrity and neuronal migration during cortical development. Nat Commun 13:2746 (2022). PubMed: 35585091
- Kuwayama R et al. Establishment of mouse model of inherited PIGO deficiency and therapeutic potential of AAV-based gene therapy. Nat Commun 13:3107 (2022). PubMed: 35661110
- Xu B et al. Mettl3-mediated m6 A modification of Lrp2 facilitates neurogenesis through Ythdc2 and elicits antidepressant-like effects. FASEB J 36:e22392 (2022). PubMed: 35716070
- Wang W et al. KAP1 phosphorylation promotes the survival of neural stem cells after ischemia/reperfusion by maintaining the stability of PCNA. Stem Cell Res Ther 13:290 (2022). PubMed: 35799276
- Hussein D et al. A paediatric dysembryoplastic neuroepithelial tumour (DNET) with deregulated stem cell markers: a case report. Transl Pediatr 11:1040-1049 (2022). PubMed: 35800288