STON1 antibody (AA 529-620)

Catalog No. ABIN524426

The Mouse Monoclonal anti-STON1 antibody has been validated for ELISA and IF and has been independently validated. It is suitable to detect STON1 in samples from Human.

Quick Overview for STON1 antibody (AA 529-620) (ABIN524426)

Target: STON1 (Stonin 1 (STON1))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This STON1 antibody is un-conjugated

Application: ELISA, Immunofluorescence (IF)

Clone: 1F3

Product Details anti-STON1 Antibody

Binding Specificity: AA 529-620

Purpose: Mouse monoclonal antibody raised against a partial recombinant SBLF.

Sequence: SLKSVVVVQG AYVELQAFVN MASLAQRSSY AGSLRSCDNI RIHFPVPSQW IKALWTMNLQ RQKSLKAKMN RRACLGSLQE LESEPVIQVT VG

Cross-Reactivity: Human

Characteristics: Antibody Reactive Against Recombinant Protein.

Immunogen: SBLF (NP_006864, 529 a.a. ~ 620 a.a) partial recombinant protein with GST tag. MW of the GST tag alone is 26 KDa.

Isotype: IgG2a

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Restrictions: For Research Use only

Independent Validation (IF)

Validation #101553 (Immunofluorescence)

by: Lab of Prof. Dr. S.A. Kushner, Department of Psychiatry, Erasmus University Medical Center

No.: #101553

Date: 01/12/2018

Antigen: STON1

Lot Number: 08242-1F3

Method validated: Immunofluorescence

Positive Control: U-373 cell line (human glioblastoma astrocytoma)

Negative Control: secondary only

Notes:

The STON1 antibody ABIN524426 specifically labels the targeted antigen in sample in IF.

Validation Images

Immunostaining of human U-373 glioblastoma cells with ABIN524426 (red, A). Plasma membranes (green, B) and nuclei (blue, C) are visualized through overexpression of a CSPG4-GFP fusion protein and DAPI. D shows a merged image of the different channels. Z-projection of image taken at 63x

Full Methods

Primary Antibody: ABIN524426

Secondary Antibody: donkey anti-mouse A647-conjugated antibody (Sanbio, 715-605-150)

Full Protocol:

• Grow U-373 cells (human glioblastoma astrocytoma) in DMEM, high glucose, glutamax (Gibco, product number: 31966-021, lot number: 1894785 supplemented with Fetal Bovine Serum (Gibco, 10082-147, lot 1907451) and penicillin streptomycin (Gibco, 15140-122, lot 1910857), at 37°C and 5% CO₂ in 2ml on a coverslips.
• Transfect cells with aCSPG4-GFP expression plasmid using X-tremeGENE HP (Sigma) accoding to the manufacturer’s recommendations.
• Fix cells on coverslips in 4% PFA for 20min at RT.
• Wash cells 3x for 5min with PBS.
• Wash coverslips for 5min with staining solution (50mM TrisHCl pH7.4, 154mM NaCl, 0.25% w/v gelatine, 5% v/v triton x-100).
• Incubate cells with primary mouse anti-human STON1 antibodylavivirus (antibodies-online, ABIN524426, lot 08242-1F3) diluted 1:100 in staining solution ON at 4°C.
• Wash cells 3x for 5min with PBS.
• Incubate cells with secondary donkey anti-mouse A647-conjugated antibody (Sanbio, 715-605-150) diluted 1:200 in staining solution for 1.5h at RT.
• Wash cells 2x for 5min with PBS.
• DAPI counterstain.
• Mount coverslips on glass slides in Mowiol 4-88 antifade reagent (Sigma, 81381, lot BCBL1789V).
• Image acquisition with LSM 700, 63x magnification, 1024x1024 resolution.

Experimental Notes:

• ABIN524426 was used in cells overexpressing CSPG4 fused to GFP. Stonin is expected to be abundantly expressed in the cells.

• No signal was detected in the secondary antibody only control (not shown).

Handling

Buffer: In 1x PBS, pH 7.4

Handling Advice: Aliquot to avoid repeated freezing and thawing.

Storage: -20 °C

Storage Comment: Store at -20°C or lower. Aliquot to avoid repeated freezing and thawing.

Target Details for STON1

Target: STON1 (Stonin 1 (STON1))

Alternative Name: STON1

Background: Full Gene Name: stonin 1
Synonyms: DKFZp781K2462,MGC149803,MGC149804,SBLF,STN1,STNB1,stoned-b1

Gene ID: 11037

NCBI Accession: NM_006873