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

Details for Product anti-KCNQ4 Antibody No. ABIN1304779, Fournisseur: Connectez-vous pour afficher
Antigène
  • KCNQ4
  • k(v)7.4
  • kv7.4
  • DFNA2
  • DFNA2A
  • KV7.4
  • potassium voltage-gated channel subfamily Q member 4
  • potassium channel, voltage gated KQT-like subfamily Q, member 4
  • potassium voltage-gated channel, subfamily Q, member 4
  • KCNQ4
  • kcnq4
  • Kcnq4
Reactivité
Mammifères
119
73
64
1
1
1
1
1
1
1
1
Hôte
Souris
74
30
17
Clonalité (Clone)
Monoclonal ()
Conjugué
Cet anticorp KCNQ4 est non-conjugé
5
5
4
4
4
4
3
3
3
3
3
3
3
3
3
3
2
1
1
1
1
1
1
1
1
1
Application
Immunostaining (ISt), Immunohistochemistry (IHC), Western Blotting (WB)
115
74
58
50
49
38
19
18
10
2
1
1
1
Options
Fournisseur
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N° du produit (Fournisseur)
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'Independent Validation' signe
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.

Anticorps primaire ABIN654696
Anticorps secondaire goat anti-mouse IgG (H+L) antibody (ImmunoReagents, GTXMU-003-DHRPX, lot 31-118-043013)
Protocole
  • 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).
Validation Images
Western Blotting validation image for anti-Potassium Voltage-Gated Channel, KQT-Like Subfamily, Member 4 (KCNQ4) antibody (ABIN1304779) Western blot analysis of KCNQ4 subunits using ABIN1304779. Total lysates from non-tra...
Clone N43-6
Isotype IgG1
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.
Plasmids, Primers & others Plasmids, Primers & others KCNQ4 products on genomics-online (e.g. as negative or positive controls)
Antigène
Autre désignation Kv7.4/KCNQ4 (KCNQ4 Antibody Extrait)
UniProt P56696
Pathways Sensory Perception of Sound
Restrictions For Research Use only
Format Liquid
Produit citée dans: 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).

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