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Mouse monoclonal antibody to Nestin [4D11] (317-630)


Catalogue No. M-1385-100
Description Nestin is a member of the class IV intermediate filament protein family which is expressed in neuronal stem cells. The molecular weight of human Nestin as determined by SDS-PAGE mobility is about 240kDa. However the real molecular weight is considerably less than this, at 177kDa, the disparity being likely due to the highly charged region of the C-terminal segment. Nestin is relatively poorly conserved in protein sequence across species boundaries, so that the mouse and human proteins have an overall identity of only 62%. As a result antibodies to the human protein often fail to recognize the rodent homologue and vice versa. However this antibody stains both rodent and human Nestin. Antibodies to Nestin are widely used to identify neural stem cells.
Batch No. See product label
Unit size 100 uL
Antigen Partial segment (region 317-630 aa) of human Nestin expressed in E.coli
Antigen Location 317-630
Antibody Type Monoclonal
Isotype IgG
Clone 4D11
Other Names Nestin; NES;
Accession P48681 NEST_HUMAN;
Produced in Mouse, IgG1 kappa
Purity Ascites fluid
Applications Western Blotting (WB), Immunocytochemistry (IC) and Flow Cytometry. Suggested dilution for WB is 1:1,000-5,000 and 1:250-500 for IC. Use 2ug/10^6 cells for Flow Cytometry. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
Comments Ready-to-use reagent for in-vitro laboratory research use only.
Specificity This antibody is specific for the 240 kDa Nestin protein by WB on developing rat brain (P18) homogenate. A much weaker band at approx. 90 kDa may also be seen. This is suggested to be a breakdown product of the 240 kDa band.
Antibody Against Nestin
Cross-reactivity Human, Rodent
Blast URL Click here
Form Lyophilised from ascites fluid.
Appearance White powder
Reconstitution Reconstitute in sterile distilled water. Centrifuge to remove any insoluble material.
Storage After reconstitution of lyophilised antibody, aliquot and store at -20C for a higher stability. Avoid freeze-thaw cycles.
Expiry Date 12 months after purchase
Specific References Schomann T et al. (2020) Multimodal imaging of hair follicle bulge-derived stem cells in a mouse model of traumatic brain injury. Cell Tissue Res. [Epub ahead of print]. Application: IHC/IF; Species: Mouse.

Schomann T et al. (2017) Neuronal differentiation of hair-follicle-bulge-derived stem cells co-cultured with mouse cochlear modiolus explants. PLos One. 12(10):e0187183. Application: ICC/IF; Species: Mouse, Hair follicle bulge-derived neural crest-derived stem cells (HFBSCs).

Gho CG et al. (2015) Isolation, expansion and neural differentiation of stem cells from human plucked hair- a further step towards autologous nerve recovery. Cytotechnology In press. Application: IF; Species: Human, Hair follicle bulge-derived neural crest-derived stem cells (HFBSCs), Keywords: Hair follicle stem cell, Regeneration, Neural crest, Neuron, Glia, Cryopreservation
Images (click to zoom)
Mouse monoclonal antibody to Nestin [4D11] (317-630) A: Mixed cultures of neonatal rat neurons and glia stained with Mouse monoclonal antibody to Nestin [4D11] M-1385-100 (red), Chicken polyclonal antibody to vimentin C-1409-50 (green) and DNA (DAPI stain, blue). Astrocytes and neuronal stem cells stain strongly and specifically in a clearly filamentous fashion with the Nestin antibody. The filamentous staining pattern is as expected as both Nestin and Vimentin are components of 10nm filaments. Note that some cells contain Nestin, but do not stain strongly for Vimentin and so appear red. Others contain Vimentin and not Nestin and so appear green- these are likely to be fibroblastic or endothelial cells. Some cells express both proteins and so appear yellowish. The presence of Nestin indicates that the cells are developing astrocytes, neuroblasts or undifferentiated neural stem cells. B: Western blot in of developing rat brain (P18) homogenate probed with mouse monoclonal antibody to Nestin M-1385-100. A single strong band running at ~240kDa is seen. C: Analysis of nesting expression in human neuroblastoma SH-SY5Y by Flow Cytometry. Fixing and Permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 1% BSA, Primary antibody: Mouse Monoclonal antibody to Nestin (cat # M-1385-100, 2μg per ~10^6 cells) for 30 minutes at room temperature, Secondary antibody: Goat anti-mouse PE labeled secondary antibody (1:100 fold dilution) with incubation for 20 minutes in dark at room temperature. Non-specific Control IgG, clone X63 (cat # M-1249-200) was used as negative control under same conditions (black dashed). Flow cytometry data and results were generated using Orflo MoxiflowTM instrument and protocols.
Mouse monoclonal antibody to Nestin [4D11] (317-630) Left: Detection of nestin expression in cortical neuron-glial cell culture from E20 rat by Immunocytochemistry. Cells were stained with mouse anti-nestin antibody (1:500, red) and with chicken antibody to MAP2 (C-1382-50, 1:5,000, green). Blue: DAPI nuclear dye. The nestin antibody labels developing astrocytes and neuronal stem cells in a clearly filamentous fashion, while the MAP2 antibody stains dendrites and perikarya of mature neurons. Right: Western blot analysis of tissue and cell lysates with mouse antibody to nestin (1:500, green). [1] protein standard, [2] embryonic E18 rat brain, [3] C6 rat glioma cells, and [4] SH-SY5Y human neuroblastoma cells.
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