Descriptions

 Shipping and Storage Conditions

 Ship with wet ice; Store at -20℃, valid for 12 months.

Descriptions

Product Information

Product Content

Descriptions

✅ 안전하고 무독성

• 본 제품은 유해하거나 독성이 있는 성분을 포함하지 않습니다.
• 고독성 metanol 등의 시약을 사용할 필요가 없습니다.

✅ 기존 Transfer buffer와 동등하거나 더 우수한 성능

• 본 제품은 기존의 일반적인 전이 버퍼와 비교하여 동일하거나 더 뛰어난 단백질 전이 효과를 제공합니다.

✅ Icabath 없이 빠른 단백질 전이

• 본 제품을 사용하면 icebath 없이 400mA의 일정한 전류 조건에서 15~30분 만에 단백질을 transfer할 수 있습니다.
• 기존 전이 버퍼보다 열 발생이 적어 실험 과정에서 과열로 인한 단백질 변성을 최소화할 수 있습니다.

Protocol

1. 1× 작동 용액(Working Solution) 제조 방법

1. 10x ice-bath free fast transfer buffer 100mL를 준비합니다.
2. 600mL의 탈이온수 또는 증류수를 추가하여 잘 혼합합니다.
3. 200mL의 무수 에탄올(100% ethanol)을 추가한 후 다시 잘 혼합합니다.
4. 최종 부피가 1L가 되도록 탈이온수 또는 증류수를 추가한 후 충분히 혼합하여 1× 작동 용액을 완성합니다.

2. 단백질 전이 과정

1. 샌드위치 구조를 완성합니다.
   • 배치 순서 (음극(검은색) → 양극(빨간색)):
     스펀지 → Filter → 젤 → membrane(PVDF 또는 NC) → Filter → 스펀지
2. Transfer tank에 배치하고 Ice-bath 없이 빠른 transfer buffer를 추가합니다.
3. 전류 400 mA로 설정 후 15~30분 동안 transfer를 수행합니다.
   • Transfer 시간이 30분 이상일 경우 icebath상에서 transfer해줘야 합니다. 해당 buffer는 열 발생을 최소화 시키나, 30분이 넘어가게 되면 단백질 변성에 영향을 끼칠 수 있는 온도변화가 있을 수 있습니다.
   • 젤 두께에 따른 전이 시간 조정
     • 0.75mm: transfer 시간 단축
     • 1.0mm: 기본 조건 (15~30분)
     • 1.5mm: transfer시간 연장

3. 전류 및 transfer 조건

4. 주의사항

1. 준비된 작동 용액은 가능한 한 빨리 사용하세요. 실험실용 초순수(G4701-500ML)로 희석할 것을 권장합니다.
2. 본 제품은 Tris-Gly, Bis-Tris 등 다양한 PAGE 젤 전이 시스템에 적합합니다.
3. PVDF 막은 사용 전 약 30초간 무수 에탄올(100% ethanol)에 담가 활성화하세요.
4. transfer 시간이 30분 이내일 경우 아이스 배스가 필요하지 않으나, 30분을 초과하는 경우 아이스 배스를 사용해야 합니다.
5. 전원 공급 장치의 브랜드와 모델에 따라 최대 전류 및 출력이 다를 수 있습니다.
   • 일부 전원 공급 장치는 400mA 설정 시 과부하 보호가 발생할 수 있으므로, 전류를 낮추거나 더 높은 출력의 전원을 사용해야 합니다.
6. 전이 버퍼 온도, 젤 두께, 아크릴아마이드 농도, 이온 농도, 여과지 두께, 전이 버퍼 사용 횟수 등이 전류와 전압에 영향을 줄 수 있습니다.
   • 과부하 발생 시 전류를 줄이고 전이 시간을 늘리는 방법을 고려하세요.
7. 제품의 정상적인 색상은 무색 투명이며, 만약 갈색 또는 황갈색으로 변할 경우 즉시 폐기하세요.
8. 저온 환경에서는 결정이 생길 수 있으나 이는 정상적인 현상입니다.
   • 결정이 생길 경우 약간 가열하거나, 자기 교반(Magnetic Stirring)을 사용하여 완전히 녹인 후 사용하세요.

 Shipping and Storage Conditions

 Ship with wet ice; Store at Room Temperatue, valid for 12 months.

Descriptions

제품은 1ea단위의 가격입니다. 구매는 최소 2개 단위부터 구매가 가능합니다.

Western Protein Marker I (Exposure)

8개의 high-purity recombinant proteins 와 4개의 pre-stained recombinant proteins으로 이루어져서 전기영동이나 transfer할 때부터 편리하게 확인할 수 있습니다.

Tris-Glycine gel range에서 15-154 kDa (~12, ~22, ~28, ~38, ~50, ~62, ~70, ~78, ~113, ~154 kDa)를 확인 할 수 있으며,  orange-red band at 70 kDa을 blue band를 13, 28, 140 kDa에서 확인할 수 있습니다. 거의 모든 종류의 antibodies 들에 적용 가능( chicken antibodies 예외).
Western Protein Marker I can bind the antibody to the target protein at the same time and develop color by ECL or other methods.

Western Protein Marker Ⅱ (Exposure)

The Western Protein Marker II는 7 high-purity recombinant proteins들과 3 prestained recombinant protein으로 구성되어 있습니다..

The bands display in Tris-Glycine gel range from 30-209 kDa (~30, 38, 50, 62, ~70, 78, ~105, ~113, 154, 209 kDa),

70 kDa는 red pre-stained band, 30 kDa와 105 kDa는 blue pre-stained band로 전기영동이나 transfer할 때 밴드를 확인하는데 용이하게 되어있습니다.

7개의 recombinant protein들 (38, 50, 62, 78, 113, 154, 209 kDa)은 거의 모든 항체들과 반응합니다. (chicken antibodies 제외)

Shipping and Storage Conditions

Ship with wet ice; Store at -20℃, valid for 12 months.

Descriptions

Product Information

Product Description/Introduction

ECL chemiluminescence Kit is a highly sensitive chemiluminescence kit based on luminol ECL chemiluminescence. This product can react with horseradish peroxidase (HRP) conjugated on secondary antibody to produce fluorescence and detect samples by using X-ray lithography or other appropriate fluorescence imaging equipment (fluorescence or chemiluminescence imaging system, etc.). Low concentration samples can still be detected, saving samples. Signal can still be detected after incubation with low concentrations of antibodies (primary, secondary), saving precious antibodies. Signal values are enhanced at least 10-fold compared to the Sensitive ECL Chemiluminescence Kit (G2074).

Storage and Shipping Conditions

Ship with wet ice; Store in the dark at 2-8℃, valid for 12 months.

Product Components

Assay Protocol/Procedures

1. hypersensitive ECL working solution 준비: the hypersensitive ECL solution A와 hypersensitive ECL solution B  같은 볼륨으로 섞음. 해당 솔루션은 실온에서 바로 사용 가능합니다.

2. Western 실험에서, PVDF membrane (또는 NC membrane)는 2nd Ab 반응 시키고, 몇 번의 washing과정을 거친 후에,  위에서 섞은 ECL working solution이 membrane을 덮을 정도로 넣은 후 1~2분간 반응 시킵니다.

3.  ECL working liquid를 filter paper나 absorbent paper로 흡수 시켜 제거하거나, membrane을 꺼내서 촬영을 위한 비닐 판 또는 보드판에 놓아 줍니다.

4.  pressed film에 사용한다면 develop과 fixed과정을 거칩니다.(G2019, G2023, G2024추천합니다.). 촬영을 합니다.

Note

1. Be sure to change the pipette tips during pipetting of Reagent A and B to avoid cross-contamination, which may result in the failure of active components.

2. Please wear gloves and use clean equipment such as tweezers to avoid contamination by exogenous proteins and metal ions during contact with the membrane.

3. Sodium azide inhibits HRP activity and all related reagents should be avoided.

4. Please use up the ECL working solution within one day after it has been configured, do not leave it until the next day, so as not to affect the accuracy of the experimental results.

5. After the use of each solution, please keep the bottle tightly capped and keep it away from light to prevent failure; especially solution B, which contains oxidizing agents and is easily reduced and become ineffective. In addition, liquid A should not be stored close to the inner wall of the refrigerator to prevent the precipitation of crystals due to the local temperature is too low. If crystals are precipitated, dissolve and mix in 37℃ warm water bath before use.

6. ECL chemiluminescence reagent kit selection refer to Table 1, primary antibody and secondary antibody recommended dilution ratio test data are from self-research antibody.

7. Refer to Table 2 for common problems and solutions for ECL chemiluminescent assays.

8. ECL chemiluminescence kit A/B solution are harmful to human body, must be careful when handling, pay attention to effective protection, avoid direct contact with the human body or inhalation of the respiratory tract.

9. For your safety and health, please wear safety glasses, gloves, or protective clothing.

Table 1: ECL Chemiluminescent Kit Selection Reference Table

Table2:Common problems and Solutions of ECL Chemiluminescence Detection

Descriptions

Product Information

Product Description/Introduction

Fast and efficient: the blocking time is only 5 minutes, and the signal-to-noise ratio is stronger than that of BSA, skim milk powder, casein and other traditional blocking agents.

Low background: this blocking solution does not contain serum, albumin and other proteins, ensuring a high signal-to-noise ratio.

Good compatibility: Compatible with horseradish peroxidase (HRP), alkaline phosphatase (AP) and biotin labeled secondary antibodies; No preservatives affecting the activity of HRP and AP; does not interfere with biotin-based assays as it does not contain biotin.

Easy to use: This product contains TBST and can be directly used for dilution of primary antibody.

Storage and Shipping Conditions

Ship with wet ice; Store at 2-8℃; Valid for 12 months; Store at -20℃ if not used for a long time.

Product Components

Assay Protocol/Procedures

1. Membrane blocking in Western blot

a) Wash with TBST for 1-2 minutes after completiing the transfer;

b) According to the size of the membrane, select a suitable container, add a certain volume of protein-free fast blocking solution to ensure cover the membrane completely, and place it on the pendulum or horizontal shaker for 5 minutes (see Question 3 for extended closure if high background occurs);

c) Dilute the primary antibody with protein-free rapid blocking solution, refer to the corresponding primary antibody instructions for dilutions, and make a primary antibody working solution.

d) After the sealing was completed, the protein-free sealing solution was poured out, washed with TBST for 10 seconds, and the primary antibody working solution was added for subsequent incubation.

Note: The primary antibody working solution must completely cover the surface of the PVDF or NC membrane; the main function of the protein-free rapid blocking buffer is to seal the sites on the PVDF or NC membrane that are not bound by proteins, so as to reduce the background, and an increase in the number of bands on the WB exposure may indicate that the antibody recognizes the non-specific bands, and may be replaced by other blocking buffer (milk or BSA);

2. Blocking of substrate in ELISA experiment

a) Use antigen or antibody coat ELISA plates

b) Add 300 μL of protein-free rapid blocking solution per well and incubate at 37 ℃ for 5 minutes;

c) Continue the following steps such as washing or shaking according to ELISA requirements.

3. blocking in IHC and IF experiments

a) This reagent is used as a solvent to Dissolve 3% (mass to volume ratio) BSA in IHC and IF experiments. It has stable sealing performance and excellent sealing effect in various tissues compared with BSA dissolved in PBS; For antibodies with good specificity, this reagent can be directly used as a blocking agent.

b) Add protein-free rapid blocking agent or BSA dissolved protein-free rapid blocking agent after preparation of the tissue sections,, and leave for 30 minutes at room temperature (with tissue differentiation); Must cover the sample completely to avoid high background from sample drying.

Note

1. No one blocking agent can be applied to all antigen and antibody detection systems. If antibodies unsuitable for this reagent, please replace other blocking agents such as BSA or skim milk powder.

2. Store the remaining solution at 2-8℃ to avoid pollution after use.

3. This reagent is viscous and should be pipetted slowly to ensure volume accuracy.

4. This product is recommended to be used only once, and repeated use may lead to the decrease of blocking effect; For antibodies with good specificity and high signal-to-noise ratio, the blocking solution can be reused; Do not mix the recovered with the unused blocking solution.

5. For your safety and health, please wear safety glasses, gloves, or protective clothing.

For Research Use Only!

Descriptions

Product Information

Storage and Shipping Conditions

Ship with wet ice; Store in the dark at 2-8℃; valid for 18 months.

Product Components

Preparation for The Experiment

Requires self-provide protease inhibitors. Protease inhibitors (G2006, G2007, G2008, etc.) should be added to RIPA Lysis (Strong) before use to prevent protein degradation. All references to RIPA Lysate (Strong) in the following instructions have been added with protease inhibitors.

Assay Protocol/Procedures

l For tissue samples

1. The tissue pieces are washed with pre-cooled PBS (G4202 recommended) to remove blood stains, cut into small pieces and placed in the homogenizer.

2. Add 10 times the tissue volume of RIPA lysis buffer (strong) and homogenate at low temperature (It is recommended to use high-speed tissue grinding instrument KZ-III-F and KZ-III-FP, which is independently developed and produced by Servicebio). Note that the amount of RIPA lysate (strong) can be added in a ratio of approximately 50 mg of tissue to 1 mL of lysate. If the content of tissue protein is low, the amount of lysate can be reduced to increase the protein concentration in crude extract solution.

3. Transfer the homogenate to a 1.5 mL of centrifuge tube and oscillate. Ice bath for 30 min with repeated pipetting every 10 min to ensure the complete lysis of tissue cells.

4. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.

l For adherent cell samples

1. Wash the cells with PBS for 2-3 times and aspirate the residual liquid thoroughly at the last time.

2. Aspirate RIPA Lysis Buffer (Strong) into the cell culture plate and flask at a ratio of 250 μL of cell lysate per well of 6-well plate, shake the culture plate and flask repeatedly to make the lysate fully contact with cells for 3-5 min.

3. Scrape off the cells with a cell scraper and collect into a centrifuge tube.

4. Lysis on ice for 30 min.

5. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.

l For suspension cell samples

1. Centrifuge.to collect cells.

2. Mix the cytosol with RIPA Lysis Buffer (Strong) at a ratio of 250 μL of lysate per cell well of a 6-well plate and shake.

3. Ice bath for 30 min, repeatedly pipetting every 10 min to ensure complete cell lysis.

4. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.

l For bacterial or fungal samples

1. Remove 1 mL of the suspension, centrifuge to remove the supernatant and wash once with PBS to fully remove the liquid. Vortex to disperse the bacterium as much as possible.

2. Add 100-200 μL of RIPA Lysis Buffer (Strong) and vortex gently to mix well with the lysate.

3. Ice bath for 30 min, repeatedly pipetting every 2 min to ensure complete cell lysis.

4. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.

Note

1. Tissue or cells may appear viscous when lysed. This can be achieved by repeatedly blowing with a pipettor or shaking with a vortexer until it becomes a liquid. Add a further appropriate amount of lysis solution if it remains thick.

2. This reagent does not contain protease inhibitors. Add your own protease inhibitors before use. G2006, G2007, G2008 and other related protease inhibitors of our company are recommended.

3. For your safety and health, please wear safety glasses, gloves, or protective clothing.

For Research Use Only!

Descriptions

Product Information

Storage and Shipping Conditions

Ship with wet ice; Store at 2-8℃ away from light; Valid for 12 months.

Product Components

Assay Protocol/Procedures

Requires self-provide protease inhibitors. Protease inhibitors (G2006, G2007, G2008, etc.) should be added to RIPA Lysis (Weak) before use to prevent protein degradation. All references to RIPA Lysate (Weak) in the following instructions have been added with protease inhibitors.

• For tissue samples:

1. The tissue pieces are washed with pre-cooled PBS (G4202 recommended) to remove blood stains, cut into small pieces and placed in the homogenizer.

2. Add 10 times the tissue volume of RIPA Lysis Buffer (Weak) and homogenate at low temperature (It is recommended to use high-speed tissue grinding instrument KZ-III-F and KZ-III-FP, which is independently developed and produced by Servicebio). Note that the amount of RIPA Lysis Buffer (Weak) can be added in a ratio of approximately 50 mg of tissue to 1 mL of lysate. If the content of tissue protein is low, the amount of lysate can be reduced to increase the protein concentration in crude extract solution.

3. Transfer the homogenate to a 1.5 mL of centrifuge tube and oscillate. Ice bath for 30 min with repeated pipetting every 10 min to ensure the complete lysis of tissue cells.

4. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.

• For adherent cell samples:

1. Wash the cells with PBS for 2-3 times and aspirate the residual liquid thoroughly at the last time.

2. Aspirate RIPA Lysis Buffer (Weak) into the cell culture plate and flask at a ratio of 250 μL of cell lysate per well of 6-well plate, shake the culture plate and flask repeatedly to make the lysate fully contact with cells for 3-5 min.

3. Scrape off the cells with a cell scraper and collect into a centrifuge tube.

4. Lysis on ice for 30 min.

5. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.

• For suspension cell samples:

1. Centrifuge to collect cells.

2. Mix the cytosol with RIPA Lysis Buffer (Weak) at a ratio of 250 μL of lysate per cell well of a 6-well plate and shake.

3. Ice bath for 30 min, repeatedly pipetting every 10 min to ensure complete cell lysis.

4. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.

• For bacterial or fungal samples:

1. Take 1 mL of the suspension, centrifuge to remove the supernatant and wash once with PBS to fully remove the liquid. Vortex to disperse the bacterium as much as possible.

2. Add 100-200 μL of RIPA Lysis Buffer (Weak) and vortex gently to mix well with the lysate.

3. Ice bath for 30 min, repeatedly pipetting every 2 min to ensure complete cell lysis.

4. Centrifuge at 12000 x g for 5 min and collect the supernatant as total protein solution.

Note

1. Tissue or cells may appear viscous when lysed. This can be achieved by repeatedly blowing with a pipettor or shaking with a vortexer until it is liquid. Add a further appropriate amount of lysis solution if it remains thick.

2. This reagent does not contain protease inhibitors. Add your own protease inhibitors before use. G2006, G2007, G2008 and other related protease inhibitors of our company are recommended.

3. For your safety and health, please wear safety glasses, gloves, or protective clothing.

For Research Use Only!