KCNQ4 Antikörper (C-Term)

Produktnummer ABIN3185286

Produktdetails anti-KCNQ4 Antikörper

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

Bindungsspezifität: C-Term

Reaktivität: Human, Maus

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser KCNQ4 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), ELISA

Spezifität: KCNQ4 Polyclonal Antibody detects endogenous levels of KCNQ4 protein.

Produktmerkmale: Rabbit Polyclonal to KCNQ4.

Aufreinigung: The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.

Immunogen: Synthesized peptide derived from the C-terminal region of human KCNQ4.

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: WB 1:500-1:2000, ELISA 1:5000,

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #102194 (Western Blotting)

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

No.: #102194

Datum: 11.04.2019

Antigen: Kv7.4/KCNQ4

Chargennummer: 3824901

Validierte Anwendung: Western Blotting

Positivkontrolle:

NCHO cells non transfected or transfected with each other member of the Kv7 family (Kv7.1, Kv7.2, Kv7.3 or Kv7.5)

Negativkontrolle:

Western blot analysis of KCNQ4/KV7.4 using ABIN3185286. Total lysates from non-transfected (NT) CHO cells or cells transfected with plasmids expressing human Kv7.4 (Kv7.4) cDNAs were prepared and separated as described in the protocol. GAPDH served as loading control.

Bewertung:

Passed. ABIN3185286 specifically detects KCNQ4/Kv7.4 in lysates from CHO cells expressing recombinant human KCNQ4/Kv7.4.

Validierungsbilder

Western blot analysis of KCNQ4/KV7.4 using ABIN3185286. Total lysates from non-transfected (NT) CHO cells or cells transfected with plasmids expressing human Kv7.4 (Kv7.4) cDNAs were prepared and separated as described in the protocol. GAPDH served as loading control.

Protokoll

Primärantikörper: ABIN3185286

Sekundärantikörper: goat anti-rabbit 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 1x Penicillin-Streptomycin mix (Microgem, L0022-100), at 37°C and 5% CO₂ in 60mm plates.
• Transfect CHO cells with plasmids expressing Kv7.4 with 5µg of plasmid each 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 100V.
• 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 40mA at 4°C ON.
• Block the membrane with blocking buffer (PBS, 5% milk dissolved, 1% Tween20) for 1h at RT.
• Incubation with primary
• • rabbit anti-KCNQ4/Kv7.4 antibody (antibodies-online, ABIN3185286, 3824901) diluted 1:500 in blocking buffer ON at 4°C or
  • rabbit anti-GAPDH (abcam, ab1816002, lot GR217575-62) diluted 1:5000 in blocking buffer ON at 4°C.
• Wash membrane 3x for 10min with PBS-T.
• Incubation with secondary goat anti-rabbit IgG (H+L) antibody (ImmunoReagents, GTXMU-003-DHRPX, lot 31-118-043013) diluted 1:500 (for ABIN3185286) or 1:20000 (for anti-GAPDH) 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 then develop and acquire signals.

Handhabung

Format: Liquid

Konzentration: 1 mg/mL

Buffer: Liquid in PBS containing 50 % glycerol, 0.5 % BSA and 0.02 % sodium azide.

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handhabung: Avoid repeated freeze/thaw cycles.

Lagerung: -20 °C

Informationen zur Lagerung: Store at -20°C.

Details zu KCNQ4

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

Andere Bezeichnung: KCNQ4 (KCNQ4 Produkte)

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

Molekulargewicht: 80 kDa

Gen-ID: 9132

UniProt: P56696

Pathways: Sensory Perception of Sound