KCNQ4 anticorps

N° du produit ABIN1304779

Détail du produit anti-KCNQ4 anticorps

Antigène: KCNQ4 (Potassium Voltage-Gated Channel, KQT-Like Subfamily, Member 4 (KCNQ4))

Reactivité: Mammifères

Hôte: Souris

Clonalité: Monoclonal

Conjugué: Cet anticorp KCNQ4 est non-conjugé

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunostaining (ISt)

Attributs du produit: K+ channels

Purification: Purified: Hybridoma in vivo ascites fluid purified by SAS (Saturated Ammonium Sulfate) precipitation followed by anion exchange chromatography. Purified mAbs are >90% specific antibody at 1mg/mL.

Clone: N43-6

Isotype: IgG1

Information d'application

Restrictions: For Research Use only

Independent Validation (WB)

Validation #102037 (Western Blotting)

by: Dipartimento di Medicina e Scienze della Salute, Università degli Studi del Molise, Campobasso

No.: #102037

Date: 31.10.2017

Antigène: Kv7.4/KCNQ4

Numéro du lot: 440-2HK-65b

Application validée: Western Blotting

Contrôle positif:

CHO cells transfected with a plasmid containing the cDNA for the longest isoform of the human Kv7.4/KCNQ4 channel

Contrôle négative:

Non-transfected CHO cells (no Kv7 channel expression)

Conclusion:

Passed. ABIN1304779 specifically detects KCNQ4/Kv7.4 in lysates from CHO cells expressing human KCNQ4/Kv7.4.

Validation Images

Western blot analysis of KCNQ4 subunits using ABIN1304779. Total lysates from non-transfected (NT) CHO cells or cells transfected with plasmids expressing human Kv7.1 (Q1; NM_019842), Kv7.2 (Q2; NM_004518.4), Kv7.3 (Q3; NM_004519.3), Kv7.4 (Q4; NM_004700.3), or Kv7.5 (Q5; NM_019842) cDNAs were prepared and separated as described in the protocol. Alpha-tubulin served as loading control.

Protocole

Anticorps primaire: ABIN654696

Anticorps secondaire: goat anti-mouse IgG (H+L) antibody (ImmunoReagents, GTXMU-003-DHRPX, lot 31-118-043013)

Full Protocol:

• Grow CHO cells (Chinese Hamster Ovary cells) in DMEM medium (Microgem, L0106-500) supplemented with 10% fetal bovine serum (Microgem, S1810-500) and antibiotics (1X Penicillin-Streptomycin mix, Microgem, L0022-100), at 37°C and 5% CO₂ in 60mm plates.
• Transfect CHO cells with 5µg plasmid expressing either Kv7.1, Kv7.2, Kv7.3, Kv7.4, or Kv7.5 and 1µg of plasmid expressing EGFP as a transfection marker using Lipofectamine 2000 (Invitrogen, 11668019) following the manufacturer´s instructions.
• Lyse cells in an appropriate volume of B-buffer, containing: 150mM NaCl, 10mM Tris-HCl pH7.4, 1mM EDTA, 1% SDS and 1x protease inhibitors (Roche, 11836145001, lot 11231400).
• Determine total protein content of the lysates using a Bradford protein assay (AppliChem, A6932, lot 6J015116).
• Denature 20µg of total protein for 10min at 100°C in 1/3 volume of 4x Laemmli SDS sample buffer and subsequently separate them on a denaturing 8% SDS-PAGE gel in Mini-PROTEAN Electrophoresis Cells (Bio-Rad) for 2-3h at 100 V.
• Transfer proteins onto PVDF filter (Bio-Rad, 1620177, lot 28369A01) with transfer buffer (25mM Tris, 192mM glycine, 20% methanol) in a western blotting system at 4°C at 40mA ON.
• Block the membrane with blocking buffer (5% milk dissolved in PBS + 1% Tween20, PBS-T) for 1h at RT.
• Incubation with primary
• • monoclonal mouse anti-KCNQ4/Kv7.4 antibody (antibodies-online, ABIN1304779, lot 440-2HK-65b) diluted 1:1000 in blocking buffer ON at 4°C or
  • mouse anti-alpha-tubulin (Sigma, T8203, lot 024M4827V) diluted 1:5000 in blocking buffer ON at 4°C.
• Wash membrane 3x for 10min with PBS-T.
• Incubation with secondary goat anti-mouse IgG (H+L) antibody (ImmunoReagents, GTXMU-003-DHRPX, lot 31-118-043013) diluted 1:5000 in blocking buffer for 1h at RT.
• Wash membrane 3x for 10min with PBS-T.
• Treat PVDF filters with chemiluminescence solutions (ECL, Promega) for 5min at RT.
• Images were captured, stored, and analyzed with the ImageLab, version 4.1 analysis software (Bio-Rad).

Stockage

Format: Liquid

Références pour anticorps anti-KCNQ4 (ABIN1304779)

Testai, Barrese, Soldovieri, Ambrosino, Martelli, Vinciguerra, Miceli, Greenwood, Curtis, Breschi, Sisalli, Scorziello, Canduela, Grandes, Calderone, Taglialatela: "Expression and function of Kv7.4 channels in rat cardiac mitochondria: possible targets for cardioprotection." dans: Cardiovascular research, Vol. 110, Issue 1, pp. 40-50, (2016) (PubMed).

Détails sur KCNQ4

Antigène: KCNQ4 (Potassium Voltage-Gated Channel, KQT-Like Subfamily, Member 4 (KCNQ4))

Autre désignation: Kv7.4/KCNQ4 (KCNQ4 Produits)

Synonymes: anticorps KCNQ4, anticorps k(v)7.4, anticorps kv7.4, anticorps DFNA2, anticorps DFNA2A, anticorps KV7.4, anticorps potassium voltage-gated channel subfamily Q member 4, anticorps potassium channel, voltage gated KQT-like subfamily Q, member 4, anticorps potassium voltage-gated channel, subfamily Q, member 4, anticorps KCNQ4, anticorps kcnq4, anticorps Kcnq4

UniProt: P56696

Pathways: Sensory Perception of Sound