Nestin Antikörper (AA 1464-1614) (FITC)

Produktnummer ABIN1774766

Der Maus Monoklonal anti-Nestin Antikörper wird verwendet zum Nachweis von Nestin in Proben von Human. Er wurde validiert für ICC, IF und FACS. Der Antikörper wurde zudem unabhängig validiert.

Kurzübersicht für Nestin Antikörper (AA 1464-1614) (FITC) (ABIN1774766)

Target: Nestin (NES)

Reaktivität: Human

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser Nestin Antikörper ist konjugiert mit FITC

Applikation: Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FACS)

Klon: 10C2

Produktdetails anti-Nestin Antikörper (FITC)

Bindungsspezifität: AA 1464-1614

Verwendungszweck: Nestin Monoclonal Antibody [10C2] (also known as 2C1.3A11) - Neural Stem Cell Marker - Fluorescein (FITC)

Spezifität: This antibody recognises nestin, a member of the intermediate filament family required for brain and eye development.

Aufreinigung: Protein G purified

Immunogen: Human nestin protein corresponding to residues 1464-1614

Isotyp: IgG1

Anwendungsinformationen

Applikationshinweise: Optimal antibody dilution should be determined by titration.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IF)

Validierung #029824 (Immunofluorescence)

by: Ensigna Biosystems

No.: #029824

Datum: 06.11.2014

Chargennummer: I030314

Validierte Anwendung: Immunofluorescence

Positivkontrolle: PC-3 cells

Negativkontrolle: MCF-7 cells

Bewertung: Faint fluorescent signal was detected in the positive control cells. No signal was detected in the negative control cells or isotype negative control.

Validierungsbilder

Legend: Immunofluorescence. Nestin (green fluorescence) is present in positive control cells (PC3) and absent in negative control cells( MCF7). Scale bar = 50 μm, Magnification = 20X, FITC Exposure = 1.5s, Insets at 40X magnification. Details: Left panel: Micrograph image of PC3 Nestin-positive prostate adenocarcinoma cells (top) and MCF7 Nestin-negative breast adenocarcinoma cells (bottom) with 15 μg/mL of anti-Nestin-FITC Ab. Middle panel: Micrograph image of PC3 Nestin-positive prostate adenocarcinoma cells (top) and MCF7 Nestin-negative breast adenocarcinoma cells (bottom) with 10 μg/mL of anti-Nestin-FITC Ab. Right panel: Micrograph image of reagent isotype control FITC-conjugated mouse IgG1 antibody on PC3 Nestin-positive prostate adenocarcinoma cells (top) and MCF7 breast adenocarcinoma cells (bottom).

Protokoll

Primärantikörper:

• Antigen: Nestin (NES) (FITC)
• Catalog number: ABIN1774766
• Lot number: I030314
• Dilution: 1:75, 1:100

Full Protocol:

• Prostate and breast adenocarcinoma cell lines were grown directly on chamber slides, washed with 1X PBS, and fixed with 4% paraformaldehyde in 1X PBS for 15 min at room temperature (RT).
• Fixed cells were rinsed three times in PBS for 5 min each at RT.
• Cells were blocked in 1X PBS / 5% normal goat serum / 0.3% Triton X-100 for 60 min at RT.
• Cells were rinsed two times in 1X PBS.
• Cells were incubated with primary antibody diluted 1:75 and 1:100 in 1X PBS / 1% BSA / 0.3% Triton X-100 overnight at 4°C in dark conditions.
• Cells were rinsed three times in PBS for 5 min each at RT.
• Coverslips were mounted on slides with DAPI-Fluoromount-G.
• Stained cells were imaged with a Nikon Eclipse E600 microscope and Olympus DP70 camera.

Anmerkungen: The anti-Nestin antibody showed positive staining of weak intensity localized appropriately to the cell cytoplasm in a pattern consistent with intermediate filaments in positive control PC3 cells. No staining was seen in the negative control MCF7 cells or with the isotype control antibody. Although specificity of the antibody was acceptable, its sensitivity with the current protocol was questionable. It is, however, entirely possible that if this antibody is used with an appropriate amplification protocol the sensitivity would be increased.

Handhabung

Format: Liquid

Buffer: PBS 0.09 % sodium azide

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Details zu Nestin

Target: Nestin (NES)

Andere Bezeichnung: Nestin

Hintergrund: ESTM 46 antibody,FLJ 21841 antibody,FLJ21841 antibody,Intermediate filament proteinantibody,Nbla00170 antibody,Nes antibody,NEST_HUMAN antibody,Nestin antibody

Gen-ID: 10763

OMIM: 600915

UniProt: P48681