Nestin Polyclonal antibody

Nestin Polyclonal Antibody for WB, IF/ICC, IF-P, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IF/ICC, IF-P, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

2

Cat no : 29285-1-AP

Synonyms

NES

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Validation Data Gallery

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  WB analysis using 29285-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 29285-1-AP (nestin antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  WB analysis using 29285-1-AP

  U-251 cells were subjected to SDS PAGE followed by western blot with 29285-1-AP (nestin antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

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  IF Staining of human kidney using 29285-1-AP

  Immunofluorescent analysis of (4% PFA) fixed human kidney tissue using Nestin antibody (29285-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of rat brain using 29285-1-AP

  Immunofluorescent analysis of (4% PFA) fixed rat brain tissue using nestin antibody (29285-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), NeuN antibody (66836-1-Ig, Clone: 3A4C1, red).

• 

  IF Staining of rat brain using 29285-1-AP

  Immunofluorescent analysis of (4% PFA) fixed rat brain tissue using nestin antibody (29285-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), GFAP antibody (60190-1-Ig, Clone: 4B2E10, red).

• 

  IF Staining of mouse kidney using 29285-1-AP

  Immunofluorescent analysis of (4% PFA) fixed mouse kidney tissue using Nestin antibody (29285-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of U-251 using 29285-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed U-251 cells using Nestin antibody (29285-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

• 

  FC experiment of SH-SY5Y using 29285-1-AP

  1x10^6 SH-SY5Y cells were intracellularly stained with 0.4 ug Anti-Human Nestin (29285-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: SGC-7901 cells, U-251 cells, U-87 MG cells
• Positive IF-P detected in: rat brain tissue, mouse kidney tissue, human kidney tissue
• Positive IF/ICC detected in: U-251 cells
• Positive FC (Intra) detected in: SH-SY5Y cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:1000
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• IF: See 2 publications below

Product Information

29285-1-AP targets Nestin in WB, IF/ICC, IF-P, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: Nestin fusion protein Ag29865
• Full Name: nestin
• Calculated Molecular Weight: 177 kDa
• Observed Molecular Weight: 240-280 kDa
• GenBank Accession Number: NM_006617
• Gene Symbol: Nestin
• Gene ID (NCBI): 10763
• RRID: AB_2918268
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Nestin, also named as NES and Nbla00170, is a class VI intermediate filament protein expressed in stem cells of the central nervous system (CNS) but not in mature CNS cells. Its expression is used extensively as a marker for neural lineage cells derived from human embryonic stem (hES) and induced pluripotent stem (iPS) cells. Nestin is also expressed in non-neural stem cell populations such as pancreatic islet progenitors and hematopoietic progenitors. Nestin can be detected as 220-240 kDa and 240-280 kDa. Studies point toward nestin heavy posttranslational modifications as possibly serving particular functions in podocytes and in a few other particular cell types(PMID: 17210924).

Publications

Species	Application	Title
human	IF	Bioact Mater; Construction of functional neural network tissue combining CBD-NT3-modified linear-ordered collagen scaffold and TrkC-modified iPSC-derived neural stem cells for spinal cord injury repair; Authors - Zhaoping Wu
mouse,rat	IF	Brain Behav Immun; Microbial reconstitution reverses prenatal stress-induced cognitive impairment and synaptic deficits in rat offspring; Authors - Jie Chen