Determination of Total Nitrogen Content in Plants by Kjeldahl Method

Product Introduction:

Measurement significance

Detection principle: The sample is boiled with concentrated sulfuric acid to convert various nitrogen-containing organic compounds into ammonium nitrogen. After alkalization, the distilled ammonia is absorbed by boric acid and titrated with standard acid to obtain the nitrogen content in the sample.
【 Friendly Reminder 】: This product is only for scientific research and should not be used for direct clinical testing on the human body. To avoid unnecessary losses, please read the purchase instructions carefully!

Features:

1. The optimized experimental plan can be completed in 1 hour

2. High sensitivity and easy operation

3. The reagent kit provides a complete set of reagents required for detecting fructose

Common sample processing methods:

1. Serum (plasma) samples: direct detection.

2. Preparation of bacterial, cell or tissue samples: Bacteria or cultured cells: Collect bacteria or cells into centrifuge tubes, centrifuge and discard supernatant; According to the number of bacteria or cells

（104 pieces): The volume of the extraction solution (mL) should be in a ratio of 500-1000:1 (it is recommended to add 1mL of extraction solution to 5 million bacteria or cells). Bacteria or cells should be sonicated (ice bath, power of 20% or 200W, sonication for 3 seconds, interval of 10 seconds, repeated 30 times) at 8000g and centrifuged at 4 ℃ for 10 minutes. The supernatant should be taken and placed on ice for testing.

Organization: According to organizational quality（g) The volume of the extraction solution (mL) should be in a ratio of 1:5-10 (it is recommended to weigh about 0.1g of tissue,

join1mL extract) was homogenized in an ice bath. Centrifuge 8000g at 4 ℃ for 10 minutes, take the supernatant, and place it on ice for testing.

Measurement steps:

1. Sample addition

1. Except for the bedding, samples need to be added at a 45 degree angle

2. The sample volume should be accurate

3. Sample should be added to the bottom of the pipe, not to the wall of the pipe

4. No bubbles should be generated during sample addition

2. Warm bath

After adding the specimen and the conjugate, they should be immediately placed in a water bath at the specified reaction temperature.

2. The ELISA plates should not be stacked together.

To avoid evaporation, the board should be covered or placed flat in a metal wet box with wet gauze at the bottom.

After adding the substrate, the reaction time and temperature are usually not strictly required. If the room temperature is above 20 ℃, the ELISA plate can be placed in the dark on the experimental bench for occasional observation. When the control tube develops color appropriately, the enzyme reaction can be terminated.

3. Washing

1. Washing is not a reaction step in the ELISA process, but it is the key to determining the success or failure of the experiment.

2. The purpose is to wash away substances in the reaction solution that have not bound to the solid-phase antigen or antibody, as well as interfering substances that have non-specific adsorption on the solid-phase carrier during the reaction process.

4. Reading board

1. The color of the negative control is extremely light, and visual colorimetry can generally be used in qualitative testing.

If the results are measured using an ELISA reader, the accuracy depends on the flatness and transparency of the ELISA plate bottom, the quality of the ELISA reader, and the algorithm of the software.

Notes:

① The order of adding reagents should be consistent to ensure that the incubation time for all reaction plate wells is the same.

② Use clean plastic containers to prepare detergent.

③ Perform incubation operations according to the time, amount, and sequence indicated in the instructions.

④ Substrate A should evaporate and avoid opening the lid for a long time. Substrate B is sensitive to light and should be avoided from prolonged exposure to light. Avoid contact with hands, toxic. After the experiment is completed, the OD value should be read immediately.

⑤ When washing the enzyme-linked immunosorbent assay (ELISA) plate, it should be thoroughly dried. Do not directly put the absorbent paper into the ELISA reaction well to absorb water.

⑥ Use disposable suction tips to avoid cross contamination, and avoid using sample dispensers with metal parts when suctioning termination solution and substrates A and B.

⑦ The Flat noodles not used in the experiment shall be immediately put back into the packaging bag and sealed for storage to avoid deterioration.

⑧ Reagents should be stored according to the label instructions and returned to room temperature before use. The diluted standard should be discarded and cannot be stored.

⑨ Other unused reagents should be packaged or covered. Do not mix reagents of different batches. Use before shelf life.

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