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Key Documents

AB5719

Sigma-Aldrich

Anti-KCa3.1 Antibody

Chemicon®, from rabbit

Synonyme(s) :

Intermediate conductance Ca2+-activated K+ Channel 4, SK4, IKCa1, KCNN4

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About This Item

Code UNSPSC :
12352203
eCl@ss :
32160702
Nomenclature NACRES :
NA.41

Source biologique

rabbit

Niveau de qualité

Forme d'anticorps

affinity purified immunoglobulin

Type de produit anticorps

primary antibodies

Clone

polyclonal

Produit purifié par

affinity chromatography

Espèces réactives

human, mouse

Fabricant/nom de marque

Chemicon®

Technique(s)

western blot: suitable

Numéro d'accès NCBI

Numéro d'accès UniProt

Conditions d'expédition

wet ice

Modification post-traductionnelle de la cible

unmodified

Informations sur le gène

human ... KCNN4(3783)

Spécificité

KCa3.1 (Intermediate conductance Ca2+-activated K+ Channel4, IKCa1, SK4, KCNN4)

Immunogène

Synthetic peptide from the C-terminal area of rat KCa3.1 (Accession Q9QYW1).

Application

Detect KCa3.1 using this Anti-KCa3.1 Antibody validated for use in WB.
Research Category
Neuroscience
Research Sub Category
Ion Channels & Transporters
Western blot: 1:300 using ECL on KCa3.1 transfected HEK-293 and K562 cells.

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).

Forme physique

Affinity purified immunoglobulin. Lyophilized from phosphate buffered saline, pH 7.4, containing 1% BSA, and 0.025% sodium azide as a preservative. Reconstitute with 50 μL of sterile deionized water. Centrifuge antibody preparation before use (10,000 xg for 5 min).

Stockage et stabilité

Maintain lyophilized material at -20°C for up to 12 months after date of receipt. After reconstitution maintain at -20°C in undiluted aliquots for up to 6 months. Avoid repeated freeze/thaw cycles.

Remarque sur l'analyse

Control
Included free of charge with the antibody is 40 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 2 μg of peptide with 1 μg of antibody for one hour at room temperature. Optimal concentrations must be determined by the end user.

Autres remarques

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Informations légales

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Clause de non-responsabilité

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.

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Mentions de danger

Conseils de prudence

Classification des risques

Aquatic Chronic 3

Code de la classe de stockage

11 - Combustible Solids

Classe de danger pour l'eau (WGK)

WGK 3


Certificats d'analyse (COA)

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