추천 제품
생물학적 소스
rabbit
Quality Level
항체 형태
affinity purified immunoglobulin
항체 생산 유형
primary antibodies
클론
polyclonal
정제법
affinity chromatography
종 반응성
rat
제조업체/상표
Chemicon®
기술
western blot: suitable
NCBI 수납 번호
UniProt 수납 번호
배송 상태
wet ice
타겟 번역 후 변형
unmodified
유전자 정보
rat ... Kcnh1(65198)
특이성
Kv10.1, EAG-1 (Ether-a-go-go K+ channel 1, KCNH1)
면역원
Synthetic peptide from the C-terminus of rat Kv10.1 (Accession Q63472). The immunogen sequence is identical in mouse and 14/16 residues identical in human.
애플리케이션
Detect Potassium Channel EAG-1 using this Anti-Potassium Channel EAG-1 Antibody validated for use in WB.
Research Category
Neuroscience
Neuroscience
Research Sub Category
Ion Channels & Transporters
Ion Channels & Transporters
Western blot: 1:200 using ECL on rat brain lysate and transfected cells HEK-Kv10.1.
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).
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).
물리적 형태
Affinity purified immunoglobulin. Lyophilized from phosphate buffered saline, pH 7.4, containing 1% BSA, and 0.025% sodium azide as a preservative. Reconstitute with 200 μL of sterile deionized water. Centrifuge antibody preparation before use (10,000 xg for 5 min).
저장 및 안정성
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.
분석 메모
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 1 μg of peptide with 1 μg of antibody for one hour at room temperature. Optimal concentrations must be determined by the end user.
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 1 μg of peptide with 1 μg of antibody for one hour at room temperature. Optimal concentrations must be determined by the end user.
기타 정보
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
법적 정보
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
면책조항
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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Hazard Classifications
Aquatic Chronic 3
Storage Class Code
11 - Combustible Solids
WGK
WGK 3
시험 성적서(COA)
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