Calretinin antibody

Catalog No. ABIN2855596

Product Details anti-Calretinin Antibody

Target: Calretinin (CALB2) (Calbindin 2 (CALB2))

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Calretinin antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunofluorescence (IF), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunocytochemistry (ICC)

Cross-Reactivity: Human, Mouse, Rat

Characteristics: Rabbit Polyclonal antibody to Calretinin (calbindin 2)
Calretinin antibody

Purification: Purified by antigen-affinity chromatography.

Immunogen: Recombinant protein encompassing a sequence within the center region of human Calretinin. The exact sequence is proprietary.

Isotype: IgG

Application Details

Application Notes: WB: 1:500-1:10000. ICC/IF: 1:100-1:1000. IHC-P: 1:100-1:1000. IHC-Fr: 1:100-1:1000. Optimal dilutions/concentrations should be determined by the researcher. Not tested in other applications.

Comment:

Positive Control: A431 , Mouse brain , human CALB2-transfected 293T cells , rat brain

Validation: Overexpression

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #103867 (Immunohistochemistry)

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

No.: #103867

Date: 05/29/2019

Antigen: Calretinin

Lot Number: 39876

Method validated: Immunohistochemistry

Positive Control:

Adult (24 months) mouse and dog brain

Negative Control:

We incubated slices overnight with the blocking solution only and then processed them with the secondary antibody.

Notes:

ABIN2855596 works in IHC-P with microwave antigen retrieval pretreatment at an optimal concentration of 1:200.

Validation Images

Staining of dog and mouse brain sections using ABIN2855596. A-C: Specific staining of calretinin-immunoreactive cortical neurons in dog brain sections at 20x, 40x, and 63x magnification. D. Staining of calretinin-immunoreactive cortical neurons in mouse brain sections at 40x magnification.

Full Methods

Primary Antibody: ABIN2855596

Secondary Antibody: goat anti-rabbit AF488-conjugated antibody (Invitrogen by Thermo Fisher Scientific, A11034, lot 1971418)

Full Protocol:

• Obtain dog samples after autopsy and immersion-fix in buffered formaldehyde for 16h.
• Perfuse mice with paraformaldehyde 4% in 0.1M phosphate buffer pH 7.4 and post-fix in the same fixative for an additional 2h at RT.
• Wash, dehydrate, and embed samples in paraffin wax.
• Cut with a microtome into 6µm sections and mount on glass slides.
• After paraffin removal, process sections by microwave antigen retrieval for 10min (95-100°C) in 10mM sodium citrate buffer (pH 6.0).
• Let sections cool for 10min.
• Wash sections under running water for 5-10min.
• Wash sections 2x 5min with distilled water.
• Wash sections 5min with 0.01M PBS.
• Incubate sections in PBS containing 1% albumin from chicken egg white (Sigma, A5378) for 1h at RT to block non-specific binding sites.
• Incubate sections with primary rabbit anti-Calretinin antibody (antibodies-online, ABIN2855596, lot 39876) diluted 1:200 in PBS-BSA-PLL at 4°C.
• Wash sections 3x 5min with 0.01M PBS.
• Incubate sections with secondary goat anti-rabbit AF488-conjugated antibody (Invitrogen by Thermo Fisher Scientific, A11034, lot 1971418) diluted 1:500 in 0.1M PBS for 1h at RT.
• Wash sections 3x 5min with 0.01M PBS.
• Mount specimens in Fluoroshield (Sigma, F6182, lot MKCB0153V).
• Acquire images with a fluorescence microscope and appropriate filter settings for AF488, e.g. Leica DM 6000B fluorescence microscope equipped with the manufacturer’s FITC filter set and digital camera at magnifications of 20x, 40x, and 63x. Parameters for image acquisition (Exposure 2.2s, Gain 2.3x, Saturation 1.5, Gamma 1.10) unchanged for all images.

Experimental Notes:

For the incubation with the primary antibody ABIN2669973 1:100, 1:500, and 1:1000 dilutions were also tested.

Handling

Format: Liquid

Concentration: 0.75 mg/mL

Buffer: 0.1M Tris-Glycine ( pH 7), 10 % Glycerol, 0.01 % Thimerosal

Preservative: Thimerosal (Merthiolate)

Precaution of Use: This product contains Thimerosal (Merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4°C. For long-term storage, aliquot and store at -20°C or below. Avoid multiple freeze-thaw cycles.

References for anti-Calretinin antibody (ABIN2855596)

Bradshaw, Trossbach, Köber, Walter, Prikulis, Weggen, Korth: "Disrupted in Schizophrenia 1 regulates the processing of reelin in the perinatal cortex." in: Schizophrenia research, Vol. 215, pp. 506-513, (2021) (PubMed).

Target Details for Calretinin

Target: Calretinin (CALB2) (Calbindin 2 (CALB2))

Alternative Name: calbindin 2 (CALB2 Products)

Synonyms: CAB29 antibody, CAL2 antibody, CR antibody, calb2l antibody, wu:fq18e08 antibody, zgc:73115 antibody, calb2 antibody, wu:fq17g09 antibody, zgc:73099 antibody, calbindin 2 antibody, calbindin 2a antibody, calbindin 2b antibody, CALB2 antibody, Calb2 antibody, calb2a antibody, calb2b antibody

Background: This gene encodes an intracellular calcium-binding protein belonging to the troponin C superfamily. Members of this protein family have six EF-hand domains which bind calcium. Three alternatively spliced transcript variants that encode different proteins have been described.

Molecular Weight: 32 kDa

Gene ID: 794

UniProt: P22676