STON1 antibody (AA 529-620)

Catalog No. ABIN524426

Product Details anti-STON1 Antibody

Target: STON1 (Stonin 1 (STON1))

Binding Specificity: AA 529-620

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This STON1 antibody is un-conjugated

Application: ELISA, Immunofluorescence (IF)

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.

Clone: 1F3

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 (STON1 Products)

Synonyms: SALF antibody, SBLF antibody, STN1 antibody, STNB1 antibody, DKFZP469K176 antibody, 4921524J06Rik antibody, C86231 antibody, Salf antibody, Sblf antibody, stonin 1 antibody, STON1 antibody, ston1 antibody, Ston1 antibody

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

Gene ID: 11037

NCBI Accession: NM_006873