STON1 anticorps (AA 529-620)

N° du produit ABIN524426

L’anticorps Souris Monoclonal anti-STON1 a été validé pour ELISA et IF et a été validé indépendamment. Il convient pour détecter STON1 dans des échantillons de Humain.

Aperçu rapide pour STON1 anticorps (AA 529-620) (ABIN524426)

Antigène: STON1 (Stonin 1 (STON1))

Reactivité: Humain

Hôte: Souris

Clonalité: Monoclonal

Conjugué: Cet anticorp STON1 est non-conjugé

Application: ELISA, Immunofluorescence (IF)

Clone: 1F3

Détail du produit anti-STON1 anticorps

Épitope: AA 529-620

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

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

Réactivité croisée: Humain

Attributs du produit: Antibody Reactive Against Recombinant Protein.

Immunogène: 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

Information d'application

Indications d'application: 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: 12.01.2018

Antigène: STON1

Numéro du lot: 08242-1F3

Application validée: Immunofluorescence

Contrôle positif: U-373 cell line (human glioblastoma astrocytoma)

Contrôle négative: secondary only

Conclusion:

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

Protocole

Anticorps primaire: ABIN524426

Anticorps secondaire: 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.

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).

Stockage

Buffer: In 1x PBS, pH 7.4

Conseil sur la manipulation: Aliquot to avoid repeated freezing and thawing.

Stock: -20 °C

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

Détails sur STON1

Antigène: STON1 (Stonin 1 (STON1))

Autre désignation: STON1

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

ID gène: 11037

NCBI Accession: NM_006873