Description
Product name Sodium Citrate Buffer with Powder (0.01mol/L, pH6.0) / Sodium Citrate Buffer with Powder (0.01mol/L, pH6.0)
Product specifications 1L | 2L
reagents are stored at room temperature and sealed, and the expiry date is at least 24 months.
Product introduction After cells or tissues are fixed with paraformaldehyde, formaldehyde or other aldehyde reagents, the intracellular antigens will form aldehyde bonds and carboxymethyl bonds, which will block part of the antigenic determinants, and at the same time cross-link between the proteins to make the antigen The determinants are concealed, leading to weakening of the staining signal during immunostaining, and even some false negative staining results. Therefore, it is required that when performing immunohistochemical staining, it is necessary to perform antigen retrieval or exposure first, that is, to destroy the crosslinks formed between molecules during fixation, and restore the original spatial shape of the antigen. Thereby increasing the detection rate of antigen, reducing background staining, and improving the accuracy of detection. Sodium citrate buffer 0.01mol/L, pH 6.0 is a commonly used antigen retrieval solution, which can be used for antigen retrieval after paraffin section, frozen section and other samples are fixed with paraformaldehyde, formaldehyde or other aldehyde reagents. It can effectively remove the crosslinks between proteins caused by aldehyde fixation reagents, and fully expose the epitopes in samples such as paraffin sections, thereby greatly improving the immunostaining effect. Normally, paraffin sections need to be processed for antigen retrieval, while frozen sections need not be processed for antigen retrieval. Antigen retrieval can improve the immunostaining effect of paraffin sections, and can also improve the staining effect of frozen sections to varying degrees. When the immunostaining effect of frozen sections is not satisfactory, consider antigen retrieval. According to the 10ml antigen retrieval solution required for each film, 1L of antigen retrieval solution can be used for antigen retrieval of 100 samples. Reference for Use This product is in dry powder form. Dissolve it to 1L or 2L in deionized water or distilled water according to the product specifications before use, and it can be used.
1. For paraffin sections:
a. Deparaffinization: xylene 3 times, 3-5 min each time → absolute ethanol 2 times, 3-5 min each time → 95% ethanol 1 time, 3-5 min → 90% ethanol 1 time, 3-5min→75% ethanol once, 3-5min→wash with distilled water twice, 3-5min each time.
b. Antigen retrieval: Immerse the slices in citric acid retrieval solution, and heat at 95-100℃ for about 15 minutes (The heating time can be controlled within 10-20 minutes. The best heating time should be explored according to different samples and target proteins. ). The citric acid repair solution needs to be preheated to 95-100°C before use. You can use an ordinary water bath or a microwave oven for heating. If you use a microwave oven for heating, you need to be careful to avoid bumping and excessive evaporation of water. Then cool to room temperature in approximately 20-30 minutes. Wash 1-2 times with immunostaining washing solution for 3-5 min each time. Subsequently, subsequent immunostaining steps such as blocking can be performed.
2. For frozen sections: Wash the sections with immunostaining washing solution for 5 minutes. Soak the slices in citric acid repair solution and heat at 95-100°C for about 15 minutes (heating time can be controlled within 10-20 minutes, and the best heating time needs to be explored according to different samples and target proteins). The citric acid repair solution needs to be preheated to 95-100°C before use. You can use an ordinary water bath or a microwave oven for heating. If you use a microwave oven for heating, take care to avoid bumping and excessive evaporation of water. Then cool to room temperature in approximately 20-30 minutes. Wash 1-2 times with immunostaining washing solution for 3-5 min each time. Subsequently, subsequent immunostaining steps such as blocking can be performed.
3. For antigen retrieval of other samples, refer to the steps of paraffin section or frozen section.
Matters needing attention For your safety and health, please wear laboratory clothes and gloves for operation.