Reelin antibody (AA 3143-3461)

Catalog No. ABIN7435136

This anti-Reelin antibody is a Rabbit Polyclonal antibody detecting Reelin in WB, IHC, IP and ICC. Suitable for Mouse. Independently validated for use in Immunohistochemistry.

Quick Overview for Reelin antibody (AA 3143-3461) (ABIN7435136)

Target: Reelin (RELN)

Reactivity: Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Reelin antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), Immunocytochemistry (ICC)

Product Details anti-Reelin Antibody

Binding Specificity: AA 3143-3461

Purpose: Polyclonal Antibody to Reelin (RELN)

Specificity: The antibody is a rabbit polyclonal antibody raised against RELN. It has been selected for its ability to recognize RELN in immunohistochemical staining and western blotting.

Cross-Reactivity: Human, Rat

Purification: Antigen-specific affinity chromatography followed by Protein A affinity chromatography

Immunogen: Recombinant Reelin (RELN) corresdonding to Glu3143~Pro3461 (Accession # Q60841)

Isotype: IgG

Application Details

Application Notes: Western blotting: 0.01-5 μg/mL,Immunohistochemistry: 5-20 μg/mL,Immunocytochemistry: 5-20 μg/mL,Optimal working dilutions must be determined by end user.

Comment:

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104517 (Immunohistochemistry)

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

No.: #104517

Date: 01/24/2024

Antigen: Reelin

Lot Number: A20230619938

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 antibody (AA 3143-3461) ABIN7435136 works in for IHC-P at 1:100 concentration 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: ABIN7435136

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 sections were processed 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, sections were washed (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, ABIN7435136, lot A20230619938) diluted 1:50 and 1:100 in PBS-BSA-PLL (Sigma, P1524) 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, 1:500, in 0.1M PBS (Thermo Fisher A11037, lot # 2439060) for 1 h at RT
• Wash sections 3x 5 min with 0.01 M PBS
• Specimens were mounted in Fluoroshield (Sigma, F6182, lot # MKCB0153V)
• Images were acquired with a Leica DM 6000B fluorescence microscope with a digital camera at 40x magnification

Handling

Format: Liquid

Concentration: 0.5 mg/mL

Buffer: PBS, pH 7.4, containing 0.02 % Sodium azide, 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: 4 °C,-20 °C

Storage Comment: Store at 4°C for frequent use. Stored at -20°C in a manual defrost freezer for two year without detectable loss of activity. Avoid repeated freeze-thaw cycles.

Expiry Date: 24 months

Target Details for Reelin

Target: Reelin (RELN)

Alternative Name: RELN

Background: RL

UniProt: Q60841

Pathways: Synaptic Membrane