Reelin antibody (N-Term)

Catalog No. ABIN6264693

The Rabbit Polyclonal anti-Reelin antibody has been validated for WB, IHC and ELISA and has been independently validated. It is suitable to detect Reelin in samples from Human, Rat and Mouse.

Quick Overview for Reelin antibody (N-Term) (ABIN6264693)

Target: Reelin (RELN)

Reactivity: Human, Rat, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Reelin antibody is un-conjugated

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

Product Details anti-Reelin Antibody

Binding Specificity: N-Term

Specificity: RELN Antibody detects endogenous levels of total RELN.

Predicted Reactivity: Pig,Horse,Sheep,Rabbit,Dog,Chicken

Purification: 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.

Isotype: IgG

Application Details

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

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104518 (Immunohistochemistry)

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

No.: #104518

Date: 01/26/2024

Antigen: Reelin

Lot Number: 10M5728

Method validated: Immunohistochemistry

Positive Control:

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

Negative Control:

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

Notes:

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

Validation Images

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

Full Methods

Primary Antibody: ABIN6264693

Secondary Antibody: 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.

Handling

Format: Liquid

Concentration: 1 mg/mL

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

Preservative: Sodium azide

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

Storage: -20 °C

Storage Comment: Store at -20 °C. Stable for 12 months from date of receipt.

Expiry Date: 12 months

Target Details for Reelin

Target: Reelin (RELN)

Alternative Name: RELN

Background:

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

Molecular Weight: 388kDa

Gene ID: 5649

UniProt: P78509

Pathways: Synaptic Membrane