Reelin Antikörper (N-Term)

Produktnummer ABIN6264693

Der Kaninchen Polyklonal Anti-Reelin-Antikörper wurde für WB, IHC und ELISA validiert und wurde unabhängig validiert. Er ist geeignet, Reelin in Proben von Human, Ratte und Maus zu detektieren.

Kurzübersicht für Reelin Antikörper (N-Term) (ABIN6264693)

Target: Reelin (RELN)

Reaktivität: Human, Ratte, Maus

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser Reelin Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunohistochemistry (IHC), ELISA

Produktdetails anti-Reelin Antikörper

Bindungsspezifität: N-Term

Spezifität: RELN Antibody detects endogenous levels of total RELN.

Homologie: Pig,Horse,Sheep,Rabbit,Dog,Chicken

Aufreinigung: The antiserum was purified by peptide affinity chromatography using SulfoLink^TM Coupling Resin (Thermo Fisher Scientific).

Immunogen: A synthesized peptide derived from human RELN, corresponding to a region within N-terminal amino acids.

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: WB 1:1000-3000, IHC 1:200, ELISA(peptide) 1:20000-1:40000

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #104518 (Immunohistochemistry)

by: Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #104518

Datum: 26.01.2024

Antigen: Reelin

Chargennummer: 10M5728

Validierte Anwendung: Immunohistochemistry

Positivkontrolle:

Postnatal day 5 and adult (> 2 months) CD1 mouse brain fixed in 4% paraformaldehyde.

Negativkontrolle:

One control slice for each experimental procedure processed omitting the primary antibody; overnight incubation in diluent solution only.

Bewertung:

Passed. The Reelin antibody (AA 491-590) ABIN6264693 works in IHC-P at 1:50 concentrations with microwave antigen retrieval.

Validierungsbilder

Reelin expression in the postnatal day 5 mouse cerebellum. EGL = external granular layer of the cerebellar cortex; IGL = internal granular layer of the cerebellar cortex;PCL/ML = Purkinje cell/molecular layers of the cerebellar cortex; WM = white matter. Staining is mainly observed in the processes of the pre-migratory granule cells. Original magnifications: A=10x; B= 20x

Protokoll

Primärantikörper: ABIN6264693

Sekundärantikörper: anti-rabbit secondary antibody Alexa Fluor 594 (Thermo Fisher Scientific, A11037, lot # 2439060)

Full Protocol:

• Perfuse mice with paraformaldehyde 4% in 0.1 M phosphate buffer pH 7.4 and post-fix of samples in the same fixative for an additional 2 h at RT.
• Wash, dehydrate, and embed samples in paraffin wax.
• Wash several times with 0.01 M PBS.
• Cut brain with a microtome into 6 µm sections and mount on glass slides.
• After paraffin removal processed sections for microwave antigen retrieval for 10 minutes (95-100° C) in 10 mM sodium citrate buffer (pH 6.0). After 20 minutes of spontaneous cooling, wash sections (distilled water, 2 times for 5 minutes each; followed by PBS for 5 minutes).
• Incubate sections for 1 h at RT in a humid chamber in PBS containing 1% ovalbumin (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot 00583) to block non-specific binding sites.
• Incubate sections with primary rabbit anti-Reelin (antibodies-online, ABIN6264693, lot 10M5728) diluted 1:50, in PBS-BSA-PLL ON at RT in a humid chamber.
• Wash sections 3x 5 min with 0.01 M PBS.
• Incubate sections with anti-rabbit secondary antibody Alexa Fluor 594 (Thermo Fisher Scientific, A11037, lot # 2439060), 1:500, in 0.1M PBS, 1 hour at room temperature.
• Wash sections 3x 5 min with 0.01 M PBS.
• Mount specimens in Fluoroshield (Sigma, F6182, lot MKCB0153V).
• Acquire images Leica DM 6000B fluorescence microscope equipped with a digital camera at 40x magnification.

Handhabung

Format: Liquid

Konzentration: 1 mg/mL

Buffer: Rabbit IgG in phosphate buffered saline , pH 7.4, 150 mM NaCl, 0.02 % sodium azide and 50 % glycerol.

Konservierungsmittel: Sodium azide

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

Lagerung: -20 °C

Informationen zur Lagerung: Store at -20 °C. Stable for 12 months from date of receipt.

Haltbarkeit: 12 months

Details zu Reelin

Target: Reelin (RELN)

Andere Bezeichnung: RELN

Hintergrund:

Description: Extracellular matrix serine protease that plays a role in layering of neurons in the cerebral cortex and cerebellum. Regulates microtubule function in neurons and neuronal migration. Affects migration of sympathetic preganglionic neurons in the spinal cord, where it seems to act as a barrier to neuronal migration. Enzymatic activity is important for the modulation of cell adhesion. Binding to the extracellular domains of lipoprotein receptors VLDLR and LRP8/APOER2 induces tyrosine phosphorylation of DAB1 and modulation of TAU phosphorylation (By similarity).

Gene: RELN

Molekulargewicht: 388kDa

Gen-ID: 5649

UniProt: P78509

Pathways: Synaptic Membrane