In 2020, the invasion of a new coronavirus (referred to as 2019-nCoV) set off a war without gunpowder. In the “epidemic” of this battle, some people went forward and went to the front line. In the face of danger and mission, you did not hesitate to rush on the front line. Behind it was the support and expectation of more than one billion people. Go forward, #decoration of new crown epidemic #, we are with you! Come on in Wuhan, come on in China!
So far, the new coronavirus pneumonia is raging in China, and the situation is still grim. In response to this, Abbkine Biotechnology Co., Ltd. has developed and produced a series of biomedical products that help the research of the new crown virus, and can also provide biomedical related technical services, involving related fields of cytology and protein research. The main products include:
|Product Number||Product name||Application||Species|
|ABP56008||ACE2 Polyclonal Antibody||ELISA, WB||Human, Mouse|
|KET6013||EliKine™ Human IL-1β ELISA Kit||ELISA||Human|
|KET6018||EliKine™ Human IL-8 ELISA Kit||ELISA||Human|
|KET6019||EliKine™ Human IL-10 ELISA Kit||ELISA||Human|
|KET6006||EliKine™ Human bEGF ELISA Kit||ELISA||Human|
|KET6007||EliKine™ Human G-CSF ELISA Kit||ELISA||Human|
|KET6008||EliKine™ Human GM-CSF ELISA Kit||ELISA||Human|
|KET6011||EliKine™ Human IFN-γ ELISA Kit||ELISA||Human|
|KET6033||EliKine™ Human VEGF ELISA Kit||ELISA||Human|
|KET6016||EliKine™ Human IL-5 ELISA Kit||ELISA||Human|
|KET6014||EliKine™ Human IL-2 ELISA Kit||ELISA||Human|
|KET6010||EliKine™ Human IFN-α ELISA Kit||ELISA||Human|
The ACE of each tissue is mainly localized to capillary endothelial cells. The lung tissue has a rich vascular bed, and the ACE contained in the pulmonary capillary bed endothelial cells is located outside the cell (ACE contained in the peripheral vascular bed endothelial cells is located inside the cell), which promotes the conversion of angiotensin I to angiotensin II. Strong effect. And pulmonary circulation is the only vascular bed in the body that does not inactivate angiotensin II. Angiotensin II is the highest in blood flowing out of the pulmonary circulation. It is generally believed that the ACE of vascular endothelial cells is closely bound to the cell membrane and hardly releases ACE, while most of the ACE produced by macrophages and monocytes are released into the blood, so when the ACE is elevated, it should be considered as macrophage-monocyte Hypersecretion of the cellular system. As shown in the table above, ABP56008 ACE2 Polyclonal Antibody is an angiotensin converting enzyme 2 polyclonal antibody, which is mainly used to specifically recognize angiotensin converting enzyme 2 and is suitable for ELISA and WB experiments. ACE2 is a receptor that confirms the entry of coronavirus β gene such as 2019-nCoV and SARS-CoV into cells. Through the study of this receptor, we can detect the viral receptors of patients with neocoronavirus pneumonia for qualitative and quantitative research.
At the same time, recent lancet magazines have provided relevant confirmatory conclusions: IL1B, IL1RA, IL7, IL8, IL9, IL10, FGF, GCSF, GMCSF, IFN-gama, IP10, MCP1, MIP1A , MIP1B, PDGF, TNF-a, and VEGF concentrations were higher in healthy adults. Plasma IL5, 1L2p70, IL15, eosinophil chemokine and RANTES levels were similar in healthy adults and patients infected with 2019-nCoV. Further comparisons between ICU patients and non-ICU patients showed that IL2 and IL7 in ICU patients. The plasma concentrations of IL10, GCSF, IP10, MCP1, MIP1A and TNF-a were higher than those in non-ICU patients. Therefore, the detection of “cytokine storm” has become an important indicator for clinical diagnosis and monitoring of the development process of New Coronavirus. In response to this important point, Abbkine Biological developed and designed the ELIKINE series of detection kits, which specifically target a variety of human cytokines, including interleukins, monocyte chemotactic factors, macrophage inflammatory factors, and vascular endothelial growth Factors and so on, the detection indicators have a wide variety and a wide range of applications.
It is worth mentioning that the advantages of Abbkine’s ELIKINE kit are obvious, including the following five points:
Firstly, the use of highly efficient, sensitive and specific antibodies, which is the key to the success of ELISA detection experiments;
Secondly, quantitative accuracy and stability repeatability and reliability;
Thirdly, the solid phase carrier with good adsorption performance, low blank value and high transparency at the bottom of the pore;
Fourthly, suitable for serum, plasma, tissue homogenate, cell culture supernatant, urine and other specimen types, a wide range of applications;
The last but not the least, cost-effective, simple operation, saving experimental expenses.
An important feature of the ELIKINE series test kit product line is that it is easy to operate and can be completed with a simple “nine-step method”. The specific steps are shown in the following figure:
- Remove the pre-coated 96-well plate, add 100 μL of gradient dilution standard and diluted sample, and incubate at room temperature for 2h;
- The reaction solution in the microtiter plate was washed a few times against absorbent paper and washed three times with 1XPBST;
- Add 100 μL of rat Biotin-labeled TNF-a antibody diluted 1: 100 and incubate for 2 h at room temperature;
- Wash 1XPBST 3 times, soak for about 1min each time
- Add 100 μL of HRP-labeled bioavidin diluted 1: 100 and incubate at room temperature for 20 min;
- Wash 1XPBST 5 times, soak for 1-2min each time;
- Incubate 90μLTMB in each well and incubate at 37 ° C in the dark for 15-20 minutes;
- 50 μL stop solution per well, mix gently
- Detection According to the operation method of the enzyme-labeled instrument, check the OD value on the machine, and obtain the concentration of the measured substance through calculation.
Having said that, many customers will care, why are there no instructions on closed methods in our steps? Is this step missing from the manual?
Here I would like to tell everyone that the enzyme-labeled plates in the ELIKINE kit received by customers are completed by antibody coating and serum blocking by our staff. Therefore, after receiving the reagent kit, the customer can open the vacuum packaging of the microtiter plate to add samples for testing. The operation is simple and saves the customer a lot of time.
At the same time, the ELIKINE kit has a wide range of applications, and the types of specimens that can be used include: liquid specimens (including serum, plasma, urine, pleural and ascites fluid, cerebrospinal fluid, cell culture supernatant, etc.); culture cells; tissue specimens. There are certain requirements for the time, method, and preservation of these specimens. For different experimental periods, the time required to save samples is different. In this regard, we have summarized the following preservation methods:
|Sample retention time||preservation method|
|Detection within a week||Store at 4 ℃|
|Detection within a month||Divide according to the amount used once, and store frozen at -20 ℃ to avoid repeated freezing and thawing.|
|Detection within 6 months||Divide according to the amount used once, and store frozen at -80 ℃ to avoid repeated freezing and thawing.|
For different samples, our pretreatment methods are different. The following are the five common typical samples:
1. Serum: Whole blood samples are left at room temperature for 2 hours or 4 ° C overnight, and centrifuged at 1000 × g, and centrifuged. After 20 minutes, the supernatant can be tested. The blood collection tube should be a disposable pyrogen-free and endotoxin-free tube.
2.Plasma: An anticoagulant should be added immediately after collecting a whole blood sample. It is recommended to use EDTA-2Na, and centrifuge at 1000 × g for 15 minutes in 30 minutes, and the supernatant can be detected. Avoid using hemolytic, hyperlipidemia samples.
3.Tissue homogenization: use pre-chilled PBS for homogenization (three methods are recommended: one is manual grinding with a Dunn’s homogenizer; the other is cell disruption with ultrasonic waves; the third is repeated freeze-thaw methods The ice crystals are formed inside the cells and the cells are broken.), Centrifuge at 5000 × g for 5-10 minutes, and take the supernatant for detection.
4.Cell culture supernatant: Take the cell culture supernatant and centrifuge at 1000 × g for 20 minutes to remove impurities and cell debris. Take the supernatant for detection.
5.Other biological samples: such as solid biological metabolites, operate according to the tissue sample processing method, and the suspended solids should be removed by centrifugation.
A large number of SARS-CoV-2 attacks the human body, causing disturbances in the human immune system, which in turn harms the host. This extreme immune attack is called a “cytokine storm”, which causes a variety of cytokines in the body fluids such as TNF-α, IL-1, IL-6, IL-12, IFN-α, IFN-β, IFN The rapid and massive production of -γ is an important cause of acute respiratory distress syndrome and multiple organ failure. In response to this major clinical conclusion, Abbkine biologically assisted the research of the new crown virus, providing cytology and proteomics related products, suitable for ELISA experiments on multiple biological indicators of patients (including virus receptor proteins and cells Factor) for qualitative and quantitative. It is hoped that in the near future, it can provide valuable testing evidence for the diagnosis and treatment of patients with new coronary pneumonia, which has profound guiding significance for clinical and scientific research.