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Nestin Antibody (S1409)

Affinity Purified Rabbit Polyclonal Antibody (Pab)

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AP2020D staining Nestin in human cerebellum tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
AP2020D staining Nestin in human brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
Overlay histogram showing SH-SY5Y cells stained with AP2020d(green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP2020d, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(OH191631) at 1/200 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
Anti-Nestin Antibody (S1409) at 1:2000 dilution + U-251 MG whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 177 kDa Blocking/Dilution buffer: 5% NFDM/TBST.

产品详情
文献引用 : 1
实验流程
背景知识

Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	IF, WB, IHC-P, FC, E
Primary Accession	P48681
Reactivity	Human, Rat, Mouse
Host	Rabbit
Clonality	Polyclonal
Isotype	Rabbit IgG
Calculated MW	177439 Da
Antigen Region	1389-1416 aa

Additional Information
Gene ID	10763
Other Names	Nestin, NES
Target/Specificity	This Nestin antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 1389-1416 amino acids from human Nestin.
Dilution	IHC-P~~1:100~500 FC~~1:10~50 WB~~1:1000 IF~~1:200
Format	Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification.
Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Precautions	Nestin Antibody (S1409) is for research use only and not for use in diagnostic or therapeutic procedures.

Protein Information
Name	NES
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 (By similarity).
Tissue Location	CNS stem cells.

Research Areas

For Research Use Only. Not For Use In Diagnostic Procedures.

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

Nestin is a class VI intermediate filament protein expressed predominantly in stem cells of the neural tube but absent from virtually all differentiated CNS cells. In the CNS, nestin is downregulated upon differentiation and replaced by neurofilaments. Transient expression of nestin has been postulated as a key step committing cells to the neural differentiation pathway. Nestin expression has also been observed in pancreatic hematopoietic stem cell populations.

REFERENCES

References for protein
1.Yaworsky, P.J., et al., Dev. Biol. 205(2):309-321 (1999).
2.Dahlstrand, J., et al., J. Cell. Sci. 103 (Pt 2), 589-597 (1992).
References for SY5Y (SH-SY5Y; ATCC#CRL-2266): 1. Ross RA, et al. Coordinate morphological and biochemical interconversion of human neuroblastoma cells. J. Natl. Cancer Inst. 71: 741-749, 1983. [PubMed: 6137586]; 2. Biedler JL, et al. Multiple neurotransmitter synthesis by human neuroblastoma cell lines and clones. Cancer Res. 38: 3751-3757, 1978. [PubMed: 29704].

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¥ 1,250.00

Cat# AP2020D

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¥ 1,250.00

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Nestin Antibody (S1409)

Affinity Purified Rabbit Polyclonal Antibody (Pab)

BACKGROUND

REFERENCES

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