KCNQ4 antibody (C-Term)

Catalog No. ABIN3185286

Product Details anti-KCNQ4 Antibody

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

Binding Specificity: C-Term

Reactivity: Human, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This KCNQ4 antibody is un-conjugated

Application: Western Blotting (WB), ELISA

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

Characteristics: Rabbit Polyclonal to KCNQ4.

Purification: 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.

Isotype: IgG

Application Details

Application Notes: WB 1:500-1:2000, ELISA 1:5000,

Restrictions: For Research Use only

Independent Validation (WB)

Validation #102194 (Western Blotting)

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

No.: #102194

Date: 04/11/2019

Antigen: Kv7.4/KCNQ4

Lot Number: 3824901

Method validated: Western Blotting

Positive Control:

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

Negative Control:

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.

Notes:

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

Validation Images

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.

Full Methods

Primary Antibody: ABIN3185286

Secondary Antibody: 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.

Handling

Format: Liquid

Concentration: 1 mg/mL

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

Preservative: Sodium azide

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

Handling Advice: Avoid repeated freeze/thaw cycles.

Storage: -20 °C

Storage Comment: Store at -20°C.

Target Details for KCNQ4

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

Alternative Name: KCNQ4 (KCNQ4 Products)

Molecular Weight: 80 kDa

Gene ID: 9132

UniProt: P56696

Pathways: Sensory Perception of Sound