The immune scatter turbidity method was used to detect the serum IgG4 level. All tests were performed using a Siemens BN II automatic protein analyzer in our hospital laboratory, with a limit of 0.08 g/L.
Bn 2 automatic protein analyzer
Manufactured by Siemens
Sourced in Germany
The BN II automatic protein analyzer is a laboratory instrument designed for the quantitative determination of proteins in a variety of biological samples. It utilizes an automated, standardized process to accurately measure protein concentrations using established analytical methods.
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3 protocols using bn 2 automatic protein analyzer
1
Serum IgG4 Level Detection
2
Comprehensive Biomarker Profiling Protocol
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Abbreviations are used to describe the biomarkers, and their reference values and categories are shown in Supplementary Table 1 .
MLR-Bf testing was performed with a Sysmex XN9000 and a Lambda Antigen Tray (ELISA). IgA, IgG, IgM, C3, C4, kapp, lamb, IgE, CRP, ASO, RF, ADNaseB, and SAA testings were performed on a SIEMENS BN II automatic protein analyzer using the original matching kit (Nephelometry). White blood cells, lymphocytes, B cells (CD3−CD19+), NK cells (CD3−CD16+CD56+), CD4+ T cells/lymphocytes (CD3+CD4+), CD8+T cells/lymphocytes (CD3+CD8+), CD3+CD4+/CD3+CD8+, and CIK cells (CD3+CD56+) were tested using the BD FACSCanto II and BD Multitest 6-color TBNK kit (flow cytometry). Anti-U1-nRNP, anti-Sm, anti-SSA-60kd, anti-Ro-52-52kd, anti-SSB, anti-Scl-70, anti-PM-Scl, anti-JO-1, anti-CENOP B, anti-PCNA, ANuA, AHA, anti-RIB-P, and AMA-M2 testings were performed using a AESKU HELIOS and AESKUSLIDES ANA Hep-2 kit (IIFA). Tests for ANA, anti-dsDNA IgG, and Anti_C1q were performed by a EUROBLOT Master and the original matching kit (Euroline).
MLR-Bf testing was performed with a Sysmex XN9000 and a Lambda Antigen Tray (ELISA). IgA, IgG, IgM, C3, C4, kapp, lamb, IgE, CRP, ASO, RF, ADNaseB, and SAA testings were performed on a SIEMENS BN II automatic protein analyzer using the original matching kit (Nephelometry). White blood cells, lymphocytes, B cells (CD3−CD19+), NK cells (CD3−CD16+CD56+), CD4+ T cells/lymphocytes (CD3+CD4+), CD8+T cells/lymphocytes (CD3+CD8+), CD3+CD4+/CD3+CD8+, and CIK cells (CD3+CD56+) were tested using the BD FACSCanto II and BD Multitest 6-color TBNK kit (flow cytometry). Anti-U1-nRNP, anti-Sm, anti-SSA-60kd, anti-Ro-52-52kd, anti-SSB, anti-Scl-70, anti-PM-Scl, anti-JO-1, anti-CENOP B, anti-PCNA, ANuA, AHA, anti-RIB-P, and AMA-M2 testings were performed using a AESKU HELIOS and AESKUSLIDES ANA Hep-2 kit (IIFA). Tests for ANA, anti-dsDNA IgG, and Anti_C1q were performed by a EUROBLOT Master and the original matching kit (Euroline).
3
Salivary Proteins and Serum α2-MG in Diabetic Patients
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Unstimulated saliva (1 mL) from the diabetic and control group was collected. Briefly, at 8 a.m., the subjects were asked to rinse their mouths thoroughly with water prior to breakfast. They were then required to tilt their heads forward, and saliva was collected into a sterile container. The saliva samples were immediately frozen and stored at -20°C until further analysis. Saliva samples were centrifuged at 2000 g for 10 min at 4°C. The supernatants were used for the detection of salivary proteins using an enzyme-linked immunosorbent assay kit (RapiBio, Inco, CA, USA) following the manufacturer instructions.
For serum samples, 3 mL venous blood was collected from the ulnar vein. Next, the blood was transferred to a vacuum blood tube. After centrifugation at 2000 g for 5 min at 4°C, the samples were immediately frozen and stored at -20°C. The concentration of α2-MG was determined using a BNII automatic protein analyzer (Siemens, Munich, Germany). The level of blood glucose was detected using aDXC800 automatic biochemistry analyzer (Beckman Coulter, Brea, CA, USA) according to manufacturer instructions.
For serum samples, 3 mL venous blood was collected from the ulnar vein. Next, the blood was transferred to a vacuum blood tube. After centrifugation at 2000 g for 5 min at 4°C, the samples were immediately frozen and stored at -20°C. The concentration of α2-MG was determined using a BNII automatic protein analyzer (Siemens, Munich, Germany). The level of blood glucose was detected using aDXC800 automatic biochemistry analyzer (Beckman Coulter, Brea, CA, USA) according to manufacturer instructions.
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