Is this loading buffer suitable for all protein electrophoresis systems?

This product is suitable for various electrophoresis buffer systems used in denaturing electrophoresis.

Do I need to boil the denatured protein after adding buffer and storing it at low temperature, and then remove it? Why?

Boiling is for protein denaturation. After adding buffer and boiling, the denatured protein has completely denatured and does not need to be boiled again. After complete dissolution, the sample can be spotted.

How to quickly remove bromophenol blue that may block the strip after running the glue?

Extending the electrophoresis time can separate bromophenol blue from the bands, and for small molecular weight proteins, it is recommended to switch to a more suitable concentration separation buffer system.

What is the reason why the protein denatured by adding buffer solution is difficult to sink to the bottom and easy to float up during sampling? How to solve it?

Perhaps due to incomplete mixing of glycerol in the buffer, it is recommended to dissolve the buffer completely at room temperature or no higher than 37 ℃ before use.

Can it be used for non denaturing electrophoresis?

No, this product contains reducing agents such as DTT and cannot be used in non denaturing electrophoresis.

When extracting suspended cell proteins, can the product be directly added and boiled for denaturation?

1 x buffer can be used for boiling directly, but it should be noted that the protein loading buffer obtained after this operation cannot be used for BCA concentration measurement.

Does this product have any requirements for protein concentration? Do we need to increase the concentration of the buffer if the protein concentration is high?

This product has no specific requirements for protein concentration, it can be diluted proportionally for use.

After adding the sample buffer to the protein sample, boiling at 95 ° C for 10 minutes, and centrifuging at 14000, some samples may experience layering before being loaded. What is the reason for this?

The sample buffer needs to be thoroughly mixed before use, and the sample and sample buffer also need to be thoroughly mixed. Boiling and denaturing after thorough mixing can avoid such situations.

The protein sample turns blue after boiling in buffer solution, but turns blue purple after being left at -80 ℃. What is the reason for this? Does it affect subsequent electrophoresis?

Bromophenol blue will change from yellow to blue purple with the pH value, which does not affect the normal use of the product. Similar discoloration does not affect subsequent electrophoresis, and the experiment can continue.

The BCA concentration measured a protein concentration of 3mg/ml, but after adding buffer, no protein bands were found. What is the reason for this?

Suggest conducting comparative experiments, setting BSA as the positive control, adding buffer solution to the protein sample lysis buffer, mixing well, boiling and denaturing, and observing the band situation by electrophoresis on the sample.

Do we need to add mercaptoethanol before using this product?

No need, this product contains DTT, which acts similarly to mercaptoethanol as a reducing agent but with lower toxicity.

At what temperature is the pH value of RIPA lysis buffer measured?

The pH value of this product is measured at room temperature and 25 ℃.

Antibody & Protein

Antibody & Protein FAQs

Elabscience® has compiled a list of the most frequently asked questions (FAQs) about the application of our antibody & protein products, with the goal of providing more efficient technical support to our customers.

This product is suitable for various electrophoresis buffer systems used in denaturing electrophoresis.
Boiling is for protein denaturation. After adding buffer and boiling, the denatured protein has completely denatured and does not need to be boiled again. After complete dissolution, the sample can be spotted.
Extending the electrophoresis time can separate bromophenol blue from the bands, and for small molecular weight proteins, it is recommended to switch to a more suitable concentration separation buffer system.
Perhaps due to incomplete mixing of glycerol in the buffer, it is recommended to dissolve the buffer completely at room temperature or no higher than 37 ℃ before use.
No, this product contains reducing agents such as DTT and cannot be used in non denaturing electrophoresis.
1 x buffer can be used for boiling directly, but it should be noted that the protein loading buffer obtained after this operation cannot be used for BCA concentration measurement.
This product has no specific requirements for protein concentration, it can be diluted proportionally for use.
The sample buffer needs to be thoroughly mixed before use, and the sample and sample buffer also need to be thoroughly mixed. Boiling and denaturing after thorough mixing can avoid such situations.
Bromophenol blue will change from yellow to blue purple with the pH value, which does not affect the normal use of the product. Similar discoloration does not affect subsequent electrophoresis, and the experiment can continue.
Suggest conducting comparative experiments, setting BSA as the positive control, adding buffer solution to the protein sample lysis buffer, mixing well, boiling and denaturing, and observing the band situation by electrophoresis on the sample.
No need, this product contains DTT, which acts similarly to mercaptoethanol as a reducing agent but with lower toxicity.
The pH value of this product is measured at room temperature and 25 ℃.

All categories

Antibody & Protein

Antibody & Protein

Antibody & Protein FAQs

Q1. Is this loading buffer suitable for all protein electrophoresis systems?

Q2. Do I need to boil the denatured protein after adding buffer and storing it at low temperature, and then remove it? Why?

Q3. How to quickly remove bromophenol blue that may block the strip after running the glue?

Q4. What is the reason why the protein denatured by adding buffer solution is difficult to sink to the bottom and easy to float up during sampling? How to solve it?

Q5. Can it be used for non denaturing electrophoresis?

Q6. When extracting suspended cell proteins, can the product be directly added and boiled for denaturation?

Q7. Does this product have any requirements for protein concentration? Do we need to increase the concentration of the buffer if the protein concentration is high?

Q8. After adding the sample buffer to the protein sample, boiling at 95 ° C for 10 minutes, and centrifuging at 14000, some samples may experience layering before being loaded. What is the reason for this?

Q9. The protein sample turns blue after boiling in buffer solution, but turns blue purple after being left at -80 ℃. What is the reason for this? Does it affect subsequent electrophoresis?

Q10. The BCA concentration measured a protein concentration of 3mg/ml, but after adding buffer, no protein bands were found. What is the reason for this?

Q11. Do we need to add mercaptoethanol before using this product?

Q12. At what temperature is the pH value of RIPA lysis buffer measured?