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AB5717-200UL Anti-Kv3.3 Antibody

AB5717-200UL
200 µL  
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      RWBRbAffinity PurifiedPolyclonal Antibody
      Description
      Catalogue NumberAB5717-200UL
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionAnti-Kv3.3 Antibody
      References
      Product Information
      FormatAffinity Purified
      Control
      • Included free of charge with the antibody is 20 μg of control antigen. The stock solution of the antigen can be made up using 100 μL of sterile distilled water. For negative control, preincubate 1 μg of peptide with 1 μg of antibody for one hour at room temperature. Optimal concentrations must be determined by the end user.
      PresentationAffinity purified immunoglobulin. Lyophilized from phosphate buffered saline, pH 7.4, containing 1% BSA, and 0.05% sodium azide as a preservative. Reconstitute with 200 μL of sterile deionized water. Centrifuge antibody preparation before use (10,000 xg for 5 min).
      Quality LevelMQ100
      Applications
      ApplicationAnti-Kv3.3 Antibody detects level of Kv3.3 & has been published & validated for use in WB.
      Key Applications
      • Western Blotting
      Application NotesWestern blot: 1:200 using ECL on rat brain membranes.

      Dilutions should be made using a carrier protein such as BSA (1-3%)

      Optimal working dilutions must be determined by the end user.

      SUGGESTED WESTERN BLOT PROTOCOL

      1. Mix the samples (organ membranes: 50 μg/lane; transfected cells: 500,000 cells/lane) with sample-buffer X 2, and heat 10 min at 70°C.

      2. 5-50 μL applied to Minigel lane (0.75-1.5 mm width) and run at standard conditions. (60 mA for 2 1.5 mm Minigel gels, 1.4 h). It is suggested that you run 5-15% acrylamide (37.5:1 acrylamide:bisacrysmide) minigel (1.5 mm width) at 30 mA/gel ~1-1.5 hours.

      3. Transfer in semi-dry system under standard conditions (3 h 100 mA for two minigel gels)

      4. Stain the transferred bands with Chemicon BLOT-FastStain (Catalog Number 2076).

      5. Destain with deionized water.

      6. Block with 5% non-fat milk (Marvel or Carnation) in PBS, and 0.025 % sodium azide, overnight at 2-8°C. The non-fat milk should be dissolved freshly, centrifuged 10,000 rpm for 10 min, and filtered through glass filter (Gelman Acrodisc).

      7. Incubation with first antibody 2 h at room temperature or overnight at 4°C in blocking solution. The antibody preparation should be centrifuged before use (10,000 g 5 min.). Optimal working dilutions and incubation time will need to be determined by the end user.

      8. Wash 4 x 10 min. with PBS-0.1% tween 20. From this stage, azide should be omitted.

      9. Incubation with the secondary antibody (HRP-conjugated goat anti-rabbit antibody, for example Chemicon Catalog Number AP132P, diluted appropriately) 1 h at room temperature.

      10. Wash 4 x 10 min. with PBS-0.1% tween 20.

      11. Perform ECL with commercial kits (Chemilucent, Chemicon Catalog Number 2600).
      Biological Information
      ImmunogenSynthetic peptide from the C-terminus of rat Kv3.3 (Accession Q01956).
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostRabbit
      SpecificityKv3.3 (KCN3). Recognizes only splice variant Kv3.3-1.
      Species Reactivity
      • Rat
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThe Shaker gene family of Drosophila encodes components of voltage-gated potassium channels and is comprised of four subfamilies. Based on sequence similarity, this gene is similar to one of these subfamilies, namely the Shaw subfamily. The protein encoded by this gene belongs to the delayed rectifier class of channel proteins and is an integral membrane protein that mediates the voltage-dependent potassium ion permeability of excitable membranes.
      Gene Symbol
      • KCNC3
      • SCA13
      • K()3.3
      • Kv3.3
      • KV3.3
      • KSHIIID
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: Q14003 # This protein mediates the voltage-dependent potassium ion permeability of excitable membranes. Assuming opened or closed conformations in response to the voltage difference across the membrane, the protein forms a potassium-selective channel through which potassium ions may pass in accordance with their electrochemical gradient.
      SIZE: 757 amino acids; 80520 Da
      SUBUNIT: Heterotetramer of potassium channel proteins (By similarity).
      SUBCELLULAR LOCATION: Membrane; Multi-pass membrane protein.
      DOMAIN: SwissProt: Q14003 The segment S4 is probably the voltage-sensor and is characterized by a series of positively charged amino acids at every third position. & The tail may be important in modulation of channel activity and/or targeting of the channel to specific subcellular compartments.
      DISEASE: SwissProt: Q14003 # Defects in KCNC3 are the cause of spinocerebellar ataxia 13 (SCA13) [MIM:605259]. Spinocerebellar ataxia is a clinically and genetically heterogeneous group of autosomal dominant cerebellar ataxias (ADCA). Patients show progressive incoordination of gait and often poor coordination of hands, speech and eye movements. Spinocerebellar ataxia is caused by degeneration of the cerebellum with variable involvement of the brainstem and spinal cord. SCA13 is a slow progressing form. Age at onset is variable, ranging from childhood to late adulthood. Mental retardation can be present in some patients.
      SIMILARITY: Belongs to the potassium channel family. C (Shaw) subfamily.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsMaintain lyophilized material at -20°C for up to 12 months. After reconstitution maintain at -20°C in undiluted aliquots for up to 6 months. Avoid repeated freeze/thaw cycles.
      Packaging Information
      Material Size200 µL
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      AB5717-200UL 04053252271915