KCNQ4 antibody
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- Target See all KCNQ4 Antibodies
- KCNQ4 (Potassium Voltage-Gated Channel, KQT-Like Subfamily, Member 4 (KCNQ4))
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Reactivity
- Mammalian
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Host
- Mouse
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Clonality
- Monoclonal
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Conjugate
- This KCNQ4 antibody is un-conjugated
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Application
- Western Blotting (WB), Immunohistochemistry (IHC), Immunostaining (ISt)
- Characteristics
- 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
- Top Product
- Discover our top product KCNQ4 Primary Antibody
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- Restrictions
- For Research Use only
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- by
- Dipartimento di Medicina e Scienze della Salute, Università degli Studi del Molise, Campobasso
- No.
- #102037
- Date
- 10/31/2017
- Antigen
- Kv7.4/KCNQ4
- Lot Number
- 440-2HK-65b
- Method validated
- Western Blotting
- Positive Control
CHO cells transfected with a plasmid containing the cDNA for the longest isoform of the human Kv7.4/KCNQ4 channel
- Negative Control
Non-transfected CHO cells (no Kv7 channel expression)
- Notes
Passed. ABIN1304779 specifically detects KCNQ4/Kv7.4 in lysates from CHO cells expressing human KCNQ4/Kv7.4.
- Primary Antibody
- ABIN654696
- Secondary Antibody
- 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).
- Experimental Notes
Validation #102037 (Western Blotting)Validation ImagesWestern 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.Full Methods -
- Format
- Liquid
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Expression and function of Kv7.4 channels in rat cardiac mitochondria: possible targets for cardioprotection." in: Cardiovascular research, Vol. 110, Issue 1, pp. 40-50, (2016) (PubMed).
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Expression and function of Kv7.4 channels in rat cardiac mitochondria: possible targets for cardioprotection." in: Cardiovascular research, Vol. 110, Issue 1, pp. 40-50, (2016) (PubMed).
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- Target
- KCNQ4 (Potassium Voltage-Gated Channel, KQT-Like Subfamily, Member 4 (KCNQ4))
- Alternative Name
- Kv7.4/KCNQ4 (KCNQ4 Products)
- Synonyms
- KCNQ4 antibody, k(v)7.4 antibody, kv7.4 antibody, DFNA2 antibody, DFNA2A antibody, KV7.4 antibody, potassium voltage-gated channel subfamily Q member 4 antibody, potassium channel, voltage gated KQT-like subfamily Q, member 4 antibody, potassium voltage-gated channel, subfamily Q, member 4 antibody, KCNQ4 antibody, kcnq4 antibody, Kcnq4 antibody
- UniProt
- P56696
- Pathways
- Sensory Perception of Sound
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