Anti-Nestin antibody [3k1] - Neural Stem Cell Marker (ab6320)

Overview

  • Product nameAnti-Nestin antibody [3k1] - Neural Stem Cell MarkerSee all Nestin primary antibodies ...
  • Description
    Mouse monoclonal [3k1] to Nestin - Neural Stem Cell Marker
  • SpecificityBased on the result of immunocytochemistry on mouse and rat neural stem cells, this antibody has no reactivity with rodent Nestin.
  • Tested applicationsIHC-Fr, IHC - Wholemount, ICC, Flow Cyt, ELISA, WB, IP, ICC/IF more details
  • Species reactivity
    Reacts with: Human

    Does not react with

    Mouse, Rat
  • Immunogen

    Nestin-transfected NS0 cells transfected with a human Nestin fragment aa residues 618-1618.

  • Positive control
    • Human neural progenitors and A172 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab6320 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
IHC-Fr Use a concentration of 0.1 µg/ml. When using aldehyde-based fixative, use at a concentration of 5 µg/ml (see Abreview).
IHC - Wholemount Use at an assay dependent concentration.
ICC Use a concentration of 5 - 10 µg/ml. Tested in human neural progenitors and A172 cells. Cells were fixed with 4% paraformaldehyde and 0.15% picric acid in PBS at RT for 20 min.
Flow Cyt Use a concentration of 25 µg/ml. Fix and permeabilize cells.
ELISA Use at an assay dependent dilution.
WB Use at an assay dependent dilution. Predicted molecular weight: 200 kDa.
IP Use at an assay dependent dilution.
ICC/IF Use at an assay dependent concentration.

Target

  • FunctionRequired 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 specificityCNS stem cells.
  • Sequence similaritiesBelongs to the intermediate filament family.
  • Developmental stageUpon terminal neural differentiation, nestin is down-regulated and replaced by neurofilaments.
  • Post-translational
    modifications
    Constitutively phosphorylated. This increases during mitosis when the cytoplasmic intermediate filament network is reorganized.
  • Target information above from: UniProt accession P48681 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • ESTM 46 antibody
    • FLJ 21841 antibody
    • FLJ21841 antibody
    • Intermediate filament protein antibody
    • Nbla00170 antibody
    • Nes antibody
    • NEST_HUMAN antibody
    • Nestin antibody
    see all

Anti-Nestin antibody [3k1] - Neural Stem Cell Marker images

  • Tumorigenicity of human glioma-derived cells. Representative immunohistochemical analysis of brain tumors generated after orthotropic injection of isolated human glioblastoma cells. Immunofluorescence of mouse brains cryosections labeled with anti-human nestin antibody. A, diffuse infiltration of the injected and contralateral hemisphere (GBM 1). B, cells infiltrating the striatum ipsilateral and contralateral to the side of injection (GBM 2). Scale bar, 100 µm. GBM xenografts recapitulate the morphology of corresponding tumors in human cancer patients.
    For xenograft tumor analysis, mice were sacrificed, and cryopreserved brain sections were cut using a 10-µm cryostat. Staining with hematoxylin-eosin identified sections bearing tumors. Briefly, brain sections were mounted on slides and stained with hematoxylin for 40 seconds and then counterstained with alcoholic eosin for 30 seconds. Cryosections containing tumors were permeabilized in PBS containing 0.5% Triton X-100 and blocked in 10% normal goat serum/PBS. After incubation with ab6320 diluted 1/1,000 in NGS-PBS overnight at 4 °C, sections were washed in Tris-buffered saline. Immunopositivity was revealed using Alexa488-conjugated goat anti-mouse IgG, 1/700. Cells were counterstained with Hoechst 33342 to identify the nuclei.
  • ab6320 staining Nestin in Human glioblastoma in mouse brain by Immunohistochemistry - Wholemount.

    Samples were incubated with primary antibody (1/1000 in diluent) for 16 hours at 4°C. An HRP-conjugated goat anti-mouse polyclonal IgG was used as the secondary antibody.

References for Anti-Nestin antibody [3k1] - Neural Stem Cell Marker (ab6320)

This product has been referenced in:
  • Garner JM  et al. Constitutive activation of signal transducer and activator of transcription 3 (STAT3) and nuclear factor ?B signaling in glioblastoma cancer stem cells regulates the Notch pathway. J Biol Chem 288:26167-76 (2013). WB . Read more (PubMed: 23902772) »
  • Ramgolam K  et al. Melanoma spheroids grown under neural crest cell conditions are highly plastic migratory/invasive tumor cells endowed with immunomodulator function. PLoS One 6:e18784 (2011). Flow Cyt ; Human . Read more (PubMed: 21526207) »

See all 11 Publications for this product

Product Wall

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - Perm wash buffer
Sample Human Cell (LGE Neural Progenitors)
Specification LGE Neural Progenitors
Gating Strategy isotype population (shown in white)
Preparation Cell harvesting/tissue preparation method: 5min collagenase followed by 5mins of accutase
Sample buffer: 5% FBS wash
Username

Dr. Adam Johnson

Verified customer

Submitted Apr 18 2014

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - Permeabilization Buffer
Sample Human Cell (Differentiated Neural Stem Cells)
Specification Differentiated Neural Stem Cells
Gating Strategy Isotype negative control (white)
Preparation Cell harvesting/tissue preparation method: Typsin-EDTA Cell Dissociation
Sample buffer: PBS and 10% FBS
Username

Abcam user community

Verified customer

Submitted Jan 29 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Sample Human Cell (hair follicle culture)
Specification hair follicle culture
Permeabilization Yes - methanol
Fixative Paraformaldehyde
Username

Mr. Heiko Locher

Verified customer

Submitted Sep 23 2013

Application Immunocytochemistry
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Sample Human Cell (H9 Differentiated Stem Cells)
Specification H9 Differentiated Stem Cells
Permeabilization Yes - 0.5% Saponin Buffer
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 19 2013

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - 0.5% Saponin Buffer
Sample Human Cell (H9 Differentiated Stem Cells)
Specification H9 Differentiated Stem Cells
Gating Strategy FL3/FSC on isotype control
Preparation Cell harvesting/tissue preparation method: Single cell dissociation buffer
Sample buffer: FACS Buffer (5% FBS)
Username

Abcam user community

Verified customer

Submitted Jul 19 2013

Abreviews
Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Sample Human Cell (Embryonic Stem Cell (EB differentiated at Day 10))
Specification Embryonic Stem Cell (EB differentiated at Day 10)
Permeabilization Yes - BD Perm/Wash Buffer
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted May 21 2013

Application Flow Cytometry
Sample Human Cell (Embryonic Stem Cell (EB differentiated at Day 10))
Specification Embryonic Stem Cell (EB differentiated at Day 10)
Preparation Cell harvesting/tissue preparation method: Trypsinisation
Sample buffer: BD Perm/Wash Buffer
Fixation Paraformaldehyde
Permeabilization Yes - BD Perm/Wash Buffer
Gating Strategy Gating position dependent on isotype control population
Username

Abcam user community

Verified customer

Submitted May 15 2013

Application Immunocytochemistry
Sample Mouse Cell (adipose derived stem cell)
Specification adipose derived stem cell
Fixative Formaldehyde
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted May 03 2013

Thank you very much for your call yesterday, and I apologize for the variation of results using this antibody.

As we discussed, I'm sending two free of charge vials to Cedarlane, who will then forward the replacement vials on to you.
Read More

Application Western blot
Sample Human Cell lysate - whole cell (Primary glioblastoma cell lines)
Loading amount 25 µg
Specification Primary glioblastoma cell lines
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Aug 03 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"