Wako Pure Chemical
 
작성일 : 13-11-19
[Wako Pure Chemical] Phos-tag® Precast Gel, SuperSep Phos-tag™
Phos-tag® Precast Gel
SuperSep Phos-tag™
  
 
Phos-tag® acrylamide를 공중합(共重合, copolymerization)한 precast gel 입니다. 개봉 후 즉시 사용할 수 있습니다. 본 제품을 사용하여 인산화 단백질을 인산화 레벨에 따라 분리 할 수 있습니다. 또한 기존의 SuperSep™ 마찬가지로 중성 gel buffer를 사용하고 있기 때문에 보존성이 뛰어나 샤프한 밴드를 얻을 수 있습니다.
 
※ 본 제품을 사용하기 전에 일반적인 SDS-PAGE 에서 영동 패턴에 문제가 없는지 확인 하십시오. ( Apply 양이 적절한 지 ,
   목적 단백질이 분해되어 있지 않거나 등).  비교 데이터를 얻으려면 같은 acrylamide 농도의 SuperSep™을 추천합니다.
   ( 참조 : Examples of use # 2)
 
 
Principle
 
클릭하시면 닫힙니다.이미지 저장을 원하시면 마우스 오른쪽클릭후 '다른이름으로 저장'을 하세요
                                                                     - 그림을 클릭하면 큰 그림이 나타납니다 -
 
 
Features
  • Ready-to-use; Your precious time is saved!
  • Isolation of phosphorylated proteins depending on the phosphorylation level
  • Good isolation with sharp bands
  • Possible to store for a long time (6 months)
 
 
Examples of Use
 
(1) Time Course of Dephosphorylation                  (2) Time Course of Dephosphorylation
  클릭하시면 닫힙니다.이미지 저장을 원하시면 마우스 오른쪽클릭후 '다른이름으로 저장'을 하세요                              클릭하시면 닫힙니다.이미지 저장을 원하시면 마우스 오른쪽클릭후 '다른이름으로 저장'을 하세요
 
[Electrophoresis buffer]
 
[Electrophoresis buffer]
 Tris-glycine-SDS electrophoresis
 buffer
 Tris-glycine-SDS buffer for electrophoresis
 
 
 
 
 
 
[Electrophoresis samples]
[Electrophoresis samples]
 
 Lane 1: β-casein (AP-treated, 0 minute)
 Lane 1: Non-treated albumin
 
 Lane 2: β-casein (AP-treated, 15 minutes)
 Lane 2: Dephosphorylated albumin
 
 Lane 3: β-casein (AP-treated, 30 minutes)
 
 
 Lane 4: β-casein (AP-treated, 45 minutes)
[Electrophoresis conditions] 20 mA, 70 minutes
 
 Lane 5: β-casein (AP-treated, 60 minutes)
[Staining]   Quick CBB staining
 
 
[Decoloration]   Deionized water
 
[Electrophoresis conditions]    35 mA, 60 minutes)
 
 
[Staining]   Quick CBB staining
 Albumin (Code No. 010-17071) was dephosph
 -orylated ausing lkaline phosphatase (NIPPON
  GENE CO., LTD.,  Code No. 319-02661).
  The result confirmed dephosphorylation by
  the band shift.
 
[Decoloration]   Deionized water
 
 
 
 
 
 
 
 
 
 β-casein was dephosphorylated in time series. The results 
 confirmed separation of phosphorylated and dephosphorylated
 β-caseins. Time-dependent dephosphorylation level  was
 also confirmed.
 
 
EasySeparator, electrophoresis tank for SuperSep
SuperSep™ 전용 전기영동장치입니다. 타사의 전기영동장치에서는 제대로 작동하지 않는 경우가 있습니다.
자세한 내용은 문의 해 주시기 바랍니다.
 
This is an electrophoresis tank for SuperSep™ precast polyacrylamide gel. Attachment and removal of the gel are easy.
Turn the knob after electrophoresis, then running buffer filled inside in the tank easily falls down. So, the gel can be removed directly.
 
Features
  • Easy set-up of SuperSep™ by turning the knob.
  • Easy removal of SuperSep™ without disposing the inside running buffer after electrophoresis.
  • Electrophoresis can be performed with a minimum amount of running buffer (250 ~ 350 mL)
 
 Easy setting
클릭하시면 닫힙니다.이미지 저장을 원하시면 마우스 오른쪽클릭후 '다른이름으로 저장'을 하세요 
  Easy removing
클릭하시면 닫힙니다.이미지 저장을 원하시면 마우스 오른쪽클릭후 '다른이름으로 저장'을 하세요
 
 
FAQ
 
Q1. Which gel staining methods can we use?
A1. Any staining methods used normally such as CBB staining, negative staining, silver staining and fluorescent staining
     can be used.
 
Q2. Decoloration is poor after CBB staining.
A2. Use of a microwave oven for decoloration gives a satisfactory result.
     Method: Transfer the stained gel in 100 mL of deionized water, add a few sheets of Kimwipe rounded, and heat in a
     microwave oven for a few minutes. Change deionized water and repeat the procedure 3 or 4 times. Be careful that the
     container is hot.
 
Q3.  Can we use the product for the western blotting?
A3. Yes, provided that zinc in the gel is eliminated with EDTA to improve transcription efficiency to membranes.
     Method: Immerse the gel in a transcription buffer (25 mmol/L tris, 192 mmol/L glycine, 10% MeOH) containing
    10 mmol/L EDTA and agitate gently for 10 minutes. Repeat the procedure 3 times. Then, agitate in a transcription buffer
    (25 mmol/L tris, 192 mmol/L glycine, 10% MeOH) not containing EDTA for 10 minutes and transcribe onto PVDF
    membrane or nitrocellulose membrane. If the transcription efficiency is poor, change the condition such as increasing the
    number of EDTA treatment or increasing EDTA concentration.
 
Q4. Bands are distorted.
A4. Samples containing EDTA, inorganic salts, surfactants, etc. may ause distortion of bands and tailing. Desalt samples by
     precipitation with TCA or dialysis.
     An empty lane also causes distortion. Apply a sample buffer (x1) in the empty lane.
 
Q5. Phosphorylation and dephosphorylation of target proteins are not separated.
A5.  Perform electrophoresis of β-casein as a positive control and alkaline phosphatase-treated β-casein as a negative
     control and check the band shift. If there is a band shift, phosphorylation and non-phosphorylation of the target protein
     may not be separated at the concentration of Phos-tag™ or acrylamide concentration in this product.
 
Q6. Can we use crude extracts of cells?
A6. Yes though the Rf value may small and isolation of bands may be obscure depending on the protein targeted.
 
Q7.  How much sample should be applied to each well?
A7. 1 to 5 μg in case of purified proteins (CBB staining), 10 to 30 μg in case of tissue or cell extracts (adjust depending on
      the amount of protein expressed).
      * These are recommended amounts. Perform an ordinary SDS-PAGE or western blotting beforehand to determine an
       appropriate sample size.
 
Q8. Which molecular markers should we use?
A8. No marker recommended. The molecular weight of a marker is not reflected in this product. Therefore, E. coli-derived
    recombination protein or dephosphorylated sample is recommended as a negative control instead of a marker.
 
Q9. What complex ion is it in the product?
A9. Zinc ion.
 
Q10. How can we know whether the band shift is due to phosphorylation?
A10. Perform electrophoresis with 12.5% SuperSep™ Ace (Wako Catalog No. 199-14971), which has the same gel
       concentration, and check if the target protein is degraded.
 
 
Ordering informations
 
Catalog Number
Product Name
Size
195-16391
  SuperSep Phos-tag™ (50 μmol/L), 12.5%, 13 wells
5 gels
193-16571
  SuperSep Phos-tag™ (50 μmol/L), 12.5%, 17 wells
5 gels
193-16691
  SuperSep Phos-tag™ (50 μmol/L), 15%, 13 wells
5 gels
196-16701
  SuperSep Phos-tag™ (50 μmol/L), 15%, 17 wells
5 gels
058-07681
 EasySeparator (an electrophoresis tank for SuperSep) *
1 set
*. Invitrogen's electrophoresis tank is applicable to SuperSep by using an adjuster. Please contact us.
 
관련제품
Catalog Number
Product Name
Size
199-14971
  SuperSep Ace (50 μmol/L), 12.5%, 13 wells
10 gels
196-14981
  SuperSep Ace (50 μmol/L), 12.5%, 17 wells
10 gels
193-14991
  SuperSep Ace (50 μmol/L), 15%, 13 wells
10 gels
190-15001
  SuperSep Ace (50 μmol/L), 15%, 17 wells
10 gels
 
 
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