Recombinant protein of human M2 pyruvate kinase (PKM2)

personM2 type pyruvate kinase (PKM2) recombinant protein

Product Name:M2 type pyruvate kinase (PKM2) recombinant protein

English nameRecombinant Pyruvate kinase isozymes M2 (PKM2)

M2PK; PKM; M2-PK; PKM1; CTHBP; OIP3; PK3; PKM; TCB; THBP1; Pyruvate kinase muscle isozyme; Thyroid hormone-binding protein 1; Cytosolic thyroid hormone-binding protein; Pyruvate Kinase, Muscle

Model:CS-D233

Enzymes and kinases

tumor immunology

Species:Homo sapiens (Human)

Source: Prokaryotic expression

hostE.coli

Endotoxin level:<1.0 EU/μ g (measured by LAL method)

Subcellular localization: nucleus, cytoplasm

Predicted molecular weight:13.4kDa

Actual molecular weight:13kDa (please refer to the instruction manual for differential analysis)

Fragments and tags:Lys141~Ala248 with N-terminal His Tag

Composition of buffer solution:20mM Tris, 150mM NaCl buffer (pH 8.0, containing 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose, and Proclin300)

Characteristic: Freeze dried powder

Purity:> 97%

Isoelectric point:5.4

Application:Positive Control; Immunogen; SDS-PAGE; WB.

Specifications:10μg50μg200μg1mg5mg

Protein method/steps:

1、 Among them, carbon and hydrogen are oxidized into carbon dioxide and water, and nitrogen in proteins is converted into ammonia and combined with sulfuric acid to form sulfuric acid chloride. Then, alkaline distillation is added to release ammonia, which is absorbed by boric acid and titrated with sulfuric acid or hydrochloric acid standard solution. Multiply the consumption of acid by a conversion factor to obtain the protein content. Based on years of operational experience, the author believes that,
During the inspection process, attention should be paid to the following three steps. 1、 Control the amount of samples and reagents added.
2、 The amount of sample to be weighed depends on the level of protein in the sample. The nitrogen content in protein is relatively constant, usually determined by measuring the nitrogen content in food. Generally, solid samples are weighed at 0.2-2.0g, semi-solid samples are weighed at 2-5g, and liquid samples are weighed at 10-20ml. For samples with low nitrogen content, the weight can be increased.
3、 2. The amount of sulfuric acid added is usually 20ml, but if the sample size exceeds 5g, the amount of sulfuric acid should be increased by 5ml per gram of sample. Otherwise, digestion of the sample will not result in lower results. 3. Addition of defoamer. Food with more fat or sugar will produce a lot of foam during digestion, which will spill out of the bottle and cause nitrogen loss, so a small amount of defoamer can be added before digestion, such as liquid paraffin, silicon defoamer, etc.
4、 4. The amount of catalyst added should be suitable for copper sulfate, which is a catalyst with good effect, low price, low environmental pollution, and is an indicator for distillation and alkali addition. Potassium sulfate can accelerate the decomposition of organic matter, raising the boiling point of sulfuric acid from 34.0 ℃ to above 40.0 ℃. However, the amount added should not be too large, otherwise the high temperature will cause the decomposition of ammonium salts and result in losses. In addition to potassium sulfate, sodium sulfate also has the same effect. 5. For difficult to digest samples, a small amount of hydrogen peroxide, sodium hypochlorite, and other oxidants can be added to accelerate the oxidation of organic matter.

5、 2. Sample digestion and processing. 1. The sample must be transferred to a dry Kjeldahl flask, otherwise the sample is prone to sticking to the bottle wall and difficult to digest. When adding sulfuric acid, the powder attached to the bottle wall should be carefully washed into the bottle to fully digest the sample. Also, pay attention to rotating the Kjeldahl flask during digestion, and use condensed acid to flush down the carbon particles attached to the bottle wall to promote digestion. After adding the sample and reagent and shaking well, a small funnel should be placed at the bottle mouth, and the bottle should be tilted 45 degrees. Tilt the asbestos mesh with small holes and be careful when heating to avoid splashing. After all the samples in the bottle are carbonized and the foam stops, increase the fire power and keep the liquid in the bottle slightly boiling until the liquid is clear and transparent, and then heat it for half an hour, and the sample will be digested. 3、 Control of conditions during distillation process.

Precautions for protein experiments:

1. Sample processing:

Sampling, storage, and processing of samples should be standardized to avoid freeze-thaw cycles. Prevent sample contamination and maintain sample purity.

2. Sample separation and preparation:

Use appropriate separation methods such as SDS-PAGE or liquid chromatography. Keep the instruments and reagents clean to prevent contamination.

3. Sample labeling and standardization:

Choose appropriate protein labeling methods to ensure labeling efficiency and stability.

4. Mass spectrometry analysis:

Ensure the calibration and performance of mass spectrometers and other related equipment are in good condition, and maintain consistency in mass spectrometry analysis conditions

5. Data processing and analysis:

Peak extraction, mass spectrometry calibration, and protein quantification should be carried out carefully using professional tools. Use statistical methods to analyze data and perform multiple hypothesis correction.

6. Technical duplication and quality control:

Perform technical repetition, including positive and negative control groups. Ensure the accuracy and reproducibility of the experiment.

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