Journal: Frontiers in Immunology

Article Title: Interleukin-35 Suppresses Interleukin-9-Secreting CD4 + T Cell Activity in Patients With Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3389/fimmu.2021.645835

Figure Lengend Snippet: IL-35 level in hepatitis B-related hepatocellular carcinoma (HCC). (A) Serum IL-35 concentration was measured by ELISA in normal controls (NC, n =11), chronic hepatitis B (CHB) patients ( n =27), and hepatitis B-related HCC patients ( n =22). Significance was assessed using one-way ANOVA and SNK- q test. (B) Serum IL-35 level was compared among hepatitis B-related HCC patients in BCLC stage A ( n =7), stage B ( n =7), stage C ( n =5), and stage D ( n =3). Significance was assessed using one-way ANOVA. (C) Serum IL-35 was also compared between hepatitis B-related HCC patients with cirrhosis ( n =14) and without cirrhosis ( n =8). Significance was assessed using Student t test. (D) Serum IL-35 was also measured in hepatitis B-related HCC patients who underwent hepatic carcinectomy (n=13) or TACE ( n =9), and was compared between baseline and 2 months post therapy. The dotted line presented lower detection limit for IL-35 (15.6 pg/mL). Significance was assessed using paired t test.

Article Snippet: Cytokine expression was measured using commercial ELISA kits (CUSABIO, Wuhan, Hubei Province, China), including human IL-35 ELISA kit (Catolog No. CSB-E13126h), human IL-9 ELISA kit (Catolog No. CSB-E04642h), human IFN-γ ELISA kit (Catolog No. CSB-E04577h), and human tumor necrosis factor-α (TNF-α) ELISA kit (Catolog No. CSB-E04740h).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Interleukin-35 Suppresses Interleukin-9-Secreting CD4 + T Cell Activity in Patients With Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3389/fimmu.2021.645835

Figure Lengend Snippet: IL-9 level in hepatitis B-related hepatocellular carcinoma (HCC). (A) Serum IL-9 concentration was measured by ELISA in normal controls (NC, n =11), chronic hepatitis B (CHB) patients ( n =27), and hepatitis B-related HCC patients ( n =22). Significance was assessed using one-way ANOVA and SNK- q test. (B) Serum IL-9 level was compared among hepatitis B-related HCC patients in BCLC stage A ( n =7), stage B ( n =7), stage C ( n =5), and stage D ( n =3). Significance was assessed using one-way ANOVA. (C) Serum IL-9 was also compared between hepatitis B-related HCC patients with cirrhosis ( n =14) and without cirrhosis ( n =8). Significance was assessed using Student t test. (D) Serum IL-9 was also measured in hepatitis B-related HCC patients who underwent hepatic carcinectomy (n=13) or TACE ( n =9), and was compared between baseline and 2 months post therapy. The dotted line presented lower detection limit for IL-9 (3.9 pg/mL). Significance was assessed using paired t test.

Article Snippet: Cytokine expression was measured using commercial ELISA kits (CUSABIO, Wuhan, Hubei Province, China), including human IL-35 ELISA kit (Catolog No. CSB-E13126h), human IL-9 ELISA kit (Catolog No. CSB-E04642h), human IFN-γ ELISA kit (Catolog No. CSB-E04577h), and human tumor necrosis factor-α (TNF-α) ELISA kit (Catolog No. CSB-E04740h).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Interleukin-35 Suppresses Interleukin-9-Secreting CD4 + T Cell Activity in Patients With Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3389/fimmu.2021.645835

Figure Lengend Snippet: Influence of Th9 cells to non-specific and HBV-specific CD8 + T cells in hepatitis B-related hepatocellular carcinoma (HCC). 10 4 of peripheral CD8 + T cells from normal controls (NC, n =6), chronic hepatitis B (CHB) patients ( n =13), and hepatitis B-related HCC patients ( n =10) were co-cultured with HepG2.2.15 cells in a direct contact manner in the presence or absence of 10 4 of autologous Th9 cells or anti-IL-9 neutralization antibody (5 μg/mL). Anti-CD3/CD28 (1 μg/mL) was added for maintenance of non-specific T cell response. Supernatants were harvested 48 hours post co-culture. (A) Percentage of target HepG2.2.15 cell death was measured by LDH release, and was compared among groups in non-specific manners. Significance was assessed using one-way ANOVA and SNK- q test. (B) IFN-γ and (C) TNF-α, which secreted by non-specific CD8 + T cells, was measured by ELISA, and was compared among groups. Significance was assessed using one-way ANOVA and SNK- q test. 10 4 of peripheral CD8 + T cells from CHB patients ( n =9), and hepatitis B-related HCC patients ( n =11) were co-cultured with HepG2.2.15 cells in a direct contact manner in the presence or absence of 10 4 of autologous Th9 cells or anti-IL-9 neutralization antibody (5 μg/mL). HBV core 18-27 epitope (sequence: FLPSDFFPSV; 5 μg/mL) was added for maintenance of HBV-specific T cell response. Supernatants were harvested 48 hours post co-culture. (D) Percentage of target HepG2.2.15 cell death was measured by LDH release, and was compared among groups in HBV core-specific manners. Significance was assessed using one-way ANOVA and SNK- q test. (E) IFN-γ and (F) TNF-α, which secreted by HBV core-specific CD8 + T cells, was measured by ELISA, and was compared among groups. The dotted line presented lower detection limit for IFN-γ (1.56 pg/mL) and TNF-α (1.95 pg/mL), respectively. Significance was assessed using one-way ANOVA and SNK- q test.

Article Snippet: Cytokine expression was measured using commercial ELISA kits (CUSABIO, Wuhan, Hubei Province, China), including human IL-35 ELISA kit (Catolog No. CSB-E13126h), human IL-9 ELISA kit (Catolog No. CSB-E04642h), human IFN-γ ELISA kit (Catolog No. CSB-E04577h), and human tumor necrosis factor-α (TNF-α) ELISA kit (Catolog No. CSB-E04740h).

Techniques: Cell Culture, Neutralization, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Sequencing

Journal: Frontiers in Immunology

Article Title: Interleukin-35 Suppresses Interleukin-9-Secreting CD4 + T Cell Activity in Patients With Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3389/fimmu.2021.645835

Figure Lengend Snippet: Influence of recombinant IL-35 to non-specific and HBV-specific Th9 cells in hepatitis B-related hepatocellular carcinoma (HCC). CD4 + CXCR3 - CCR4 - CCR6 - cells were purified from peripheral blood mononuclear cells (PBMCs) of chronic hepatitis B (CHB) patients ( n =7) and hepatitis B-related HCC patients ( n =9), as well as from intrahepatic lymphocytes (IHLs) of fresh HCC specimens and non-tumor site liver specimens in HCC patients ( n =6). 10 4 of CD4 + CXCR3 - CCR4 - CCR6 - cells were stimulated with either anti-CD3/CD28 (1 μg/mL) or HBsAg (10 μg/mL) in the presence or absence of recombinant IL-35 (1 ng/mL) for 24 hours. Supernatants and cells were harvested. IL-9 level in the supernatants was measured by ELISA, while PU.1 mRNA expression in Th9 cells was quantified by real-time PCR. (A) IL-9 secretion by non-specific Th9 cells was compared among groups with or without IL-35 stimulation. Significance was assessed using one-way ANOVA and SNK- q test. (B) IL-9 secretion by HBV-specific Th9 cells was compared among groups with or without IL-35 stimulation. Significance was assessed using one-way ANOVA and SNK- q test. (C) PU.1 mRNA expression in non-specific Th9 cells was compared among groups with or without IL-35 stimulation. Significance was assessed using one-way ANOVA and SNK- q test. (D) PU.1 mRNA expression in HBV-specific Th9 cells was compared among groups with or without IL-35 stimulation. The dotted line presented lower detection limit for IL-9 (3.9 pg/mL). Significance was assessed using one-way ANOVA and SNK- q test.

Article Snippet: Cytokine expression was measured using commercial ELISA kits (CUSABIO, Wuhan, Hubei Province, China), including human IL-35 ELISA kit (Catolog No. CSB-E13126h), human IL-9 ELISA kit (Catolog No. CSB-E04642h), human IFN-γ ELISA kit (Catolog No. CSB-E04577h), and human tumor necrosis factor-α (TNF-α) ELISA kit (Catolog No. CSB-E04740h).

Techniques: Recombinant, Purification, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction

Journal: Cancer cell

Article Title: Th9 Cells Represent a Unique Subset of CD4 + T Cells Endowed with the Ability to Eradicate Advanced Tumors

doi: 10.1016/j.ccell.2018.05.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ELISA kits mouse IL-9 , eBioscience , 50-112-5217.

Techniques: Blocking Assay, Staining, Extraction, Enzyme-linked Immunosorbent Assay, Luminescence Assay, Recombinant, shRNA, Negative Control, Binding Assay, Modification, Software, Gene Expression

Journal: Central-European journal of immunology

Article Title: Targeting the serum marker interleukin 9 improves the underlying characterization and immune homeostasis in rheumatoid arthritis.

doi: 10.5114/ceji.2024.141695

Figure Lengend Snippet: Fig. 3. Interleukin 9 affected the severity of arthritis in CIA rats. A) ELISA for the detection of IL-9 levels. B) Rat arthritis score. C) Analysis of cartilage damage by saffron O-solid green staining. The descriptive statistics are pre- sented as mean ±SD. *p < 0.05 vs. the CIA group, n = 3

Article Snippet: We measured the levels of serum markers using ELISA with a specialized commercial human IL-9 ELISA Kit (CSB-E04642h, Cusabio, Wuhan, China), human Gal1 ELISA Kit (CSB-EL012882h, Cusabio), human anti-CII ELISA Kit (HB3371-Hu, Shanghai Hengyuan Biotechnology Co., Ltd, Shanghai, China), human sSR-A ELISA Kit (HB3370-Hu, Shanghai Hengyuan Biotechnology Co., Ltd), human anti-Sa ELISA Kit (ml382511V), human RF ELISA Kit (ml060678V), human ACPA ELISA Kit (ml233305V), rat IL-9 ELISA Kit (CSB-E07294r, Cusabio), rat Gal1 ELISA Kit (JL25762, Shanghai Jianglai Biotechnology Co., Ltd, Shanghai, China), rat anti-CII ELISA Kit (HB1364-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat sSR-A ELISA Kit (HB1363-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat anti-Sa ELISA Kit (ml202241V), rat RF ELISA Kit (ml059541V), and rat ACPA ELISA Kit (ml201458V).

Techniques: Enzyme-linked Immunosorbent Assay, Staining

Journal: Central-European Journal of Immunology

Article Title: Targeting the serum marker interleukin 9 improves the underlying characterization and immune homeostasis in rheumatoid arthritis

doi: 10.5114/ceji.2024.141695

Figure Lengend Snippet: Detection of serum marker levels in rheumatoid arthritis (RA) patients with different antibody types. A ) ELISA for the determination of IL-9, Gal1, anti-CII, sSR-A, anti-Sa, RF, and ACPA levels in serum of clinical samples. * p < 0.05 vs. the normal group, # p < 0.05 vs. the RA + RF negative group, & p < 0.05 vs. the RA + ACPA negative group. B ) Spearman’s analysis of the correlation of the above serum markers. The descriptive statistics are presented as mean ±SD. * p < 0.05, ** p < 0.01, **** p < 0.0001, ns. represents no significance. n = 4 in normal group, n = 10 in RA + RF negative group, n = 12 in RA + ACPA negative group, n = 13 in SNRA group

Article Snippet: We measured the levels of serum markers using ELISA with a specialized commercial human IL-9 ELISA Kit (CSB-E04642h, Cusabio, Wuhan, China), human Gal1 ELISA Kit (CSB-EL012882h, Cusabio), human anti-CII ELISA Kit (HB3371-Hu, Shanghai Hengyuan Biotechnology Co., Ltd, Shanghai, China), human sSR-A ELISA Kit (HB3370-Hu, Shanghai Hengyuan Biotechnology Co., Ltd), human anti-Sa ELISA Kit (ml382511V), human RF ELISA Kit (ml060678V), human ACPA ELISA Kit (ml233305V), rat IL-9 ELISA Kit (CSB-E07294r, Cusabio), rat Gal1 ELISA Kit (JL25762, Shanghai Jianglai Biotechnology Co., Ltd, Shanghai, China), rat anti-CII ELISA Kit (HB1364-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat sSR-A ELISA Kit (HB1363-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat anti-Sa ELISA Kit (ml202241V), rat RF ELISA Kit (ml059541V), and rat ACPA ELISA Kit (ml201458V).

Techniques: Marker, Enzyme-linked Immunosorbent Assay

Journal: Central-European Journal of Immunology

Article Title: Targeting the serum marker interleukin 9 improves the underlying characterization and immune homeostasis in rheumatoid arthritis

doi: 10.5114/ceji.2024.141695

Figure Lengend Snippet: Validation of serum marker expression in CIA rats. A ) Rat paw volume. B ) Rat arthritis score. C ) H&E staining was used to assess histopathologic damage in joints. D ) ELISA for serum markers (IL-9, Gal1, anti-CII, sSR-A, anti-a, RF, ACPA) in rats. The descriptive statistics are presented as mean ±SD. * p < 0.05, ** p < 0.01, n = 3 E ) Pearson analysis of the correlation between IL-9 and arthritis scores. The descriptive statistics are presented as mean ±SD. * p < 0.05, ** p < 0.01, n = 3

Article Snippet: We measured the levels of serum markers using ELISA with a specialized commercial human IL-9 ELISA Kit (CSB-E04642h, Cusabio, Wuhan, China), human Gal1 ELISA Kit (CSB-EL012882h, Cusabio), human anti-CII ELISA Kit (HB3371-Hu, Shanghai Hengyuan Biotechnology Co., Ltd, Shanghai, China), human sSR-A ELISA Kit (HB3370-Hu, Shanghai Hengyuan Biotechnology Co., Ltd), human anti-Sa ELISA Kit (ml382511V), human RF ELISA Kit (ml060678V), human ACPA ELISA Kit (ml233305V), rat IL-9 ELISA Kit (CSB-E07294r, Cusabio), rat Gal1 ELISA Kit (JL25762, Shanghai Jianglai Biotechnology Co., Ltd, Shanghai, China), rat anti-CII ELISA Kit (HB1364-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat sSR-A ELISA Kit (HB1363-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat anti-Sa ELISA Kit (ml202241V), rat RF ELISA Kit (ml059541V), and rat ACPA ELISA Kit (ml201458V).

Techniques: Biomarker Discovery, Marker, Expressing, Staining, Enzyme-linked Immunosorbent Assay

Journal: Central-European Journal of Immunology

Article Title: Targeting the serum marker interleukin 9 improves the underlying characterization and immune homeostasis in rheumatoid arthritis

doi: 10.5114/ceji.2024.141695

Figure Lengend Snippet: Interleukin 9 affected the severity of arthritis in CIA rats. A ) ELISA for the detection of IL-9 levels. B ) Rat arthritis score. C ) Analysis of cartilage damage by saffron O-solid green staining. The descriptive statistics are presented as mean ±SD. * p < 0.05 vs. the CIA group, n = 3

Article Snippet: We measured the levels of serum markers using ELISA with a specialized commercial human IL-9 ELISA Kit (CSB-E04642h, Cusabio, Wuhan, China), human Gal1 ELISA Kit (CSB-EL012882h, Cusabio), human anti-CII ELISA Kit (HB3371-Hu, Shanghai Hengyuan Biotechnology Co., Ltd, Shanghai, China), human sSR-A ELISA Kit (HB3370-Hu, Shanghai Hengyuan Biotechnology Co., Ltd), human anti-Sa ELISA Kit (ml382511V), human RF ELISA Kit (ml060678V), human ACPA ELISA Kit (ml233305V), rat IL-9 ELISA Kit (CSB-E07294r, Cusabio), rat Gal1 ELISA Kit (JL25762, Shanghai Jianglai Biotechnology Co., Ltd, Shanghai, China), rat anti-CII ELISA Kit (HB1364-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat sSR-A ELISA Kit (HB1363-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat anti-Sa ELISA Kit (ml202241V), rat RF ELISA Kit (ml059541V), and rat ACPA ELISA Kit (ml201458V).

Techniques: Enzyme-linked Immunosorbent Assay, Staining

Journal: Central-European Journal of Immunology

Article Title: Targeting the serum marker interleukin 9 improves the underlying characterization and immune homeostasis in rheumatoid arthritis

doi: 10.5114/ceji.2024.141695

Figure Lengend Snippet: Targeted inhibition of IL-9 improves inflammation and Treg/Th17 immune balance in CIA rats. A ) Proportion of Treg and Th17 cells infiltrated in arthritic tissues by flow cytometry assay. B ) Change in Treg/Th17 ratio. C ) RT-qPCR detection of foxp3 and IL-17A expression in arthritic tissues. D ) Determination of TNF-α, IL-1β, IL-6, and IL-17A levels in the synovial serum of CIA rats by ELISA. The descriptive statistics are presented as mean ±SD. * p < 0.05 vs. the CIA group, n = 3

Article Snippet: We measured the levels of serum markers using ELISA with a specialized commercial human IL-9 ELISA Kit (CSB-E04642h, Cusabio, Wuhan, China), human Gal1 ELISA Kit (CSB-EL012882h, Cusabio), human anti-CII ELISA Kit (HB3371-Hu, Shanghai Hengyuan Biotechnology Co., Ltd, Shanghai, China), human sSR-A ELISA Kit (HB3370-Hu, Shanghai Hengyuan Biotechnology Co., Ltd), human anti-Sa ELISA Kit (ml382511V), human RF ELISA Kit (ml060678V), human ACPA ELISA Kit (ml233305V), rat IL-9 ELISA Kit (CSB-E07294r, Cusabio), rat Gal1 ELISA Kit (JL25762, Shanghai Jianglai Biotechnology Co., Ltd, Shanghai, China), rat anti-CII ELISA Kit (HB1364-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat sSR-A ELISA Kit (HB1363-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat anti-Sa ELISA Kit (ml202241V), rat RF ELISA Kit (ml059541V), and rat ACPA ELISA Kit (ml201458V).

Techniques: Inhibition, Flow Cytometry, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Central-European Journal of Immunology

Article Title: Targeting the serum marker interleukin 9 improves the underlying characterization and immune homeostasis in rheumatoid arthritis

doi: 10.5114/ceji.2024.141695

Figure Lengend Snippet: Targeted inhibition of IL-9 regulates M2 and M1 macrophage activation. A ) Immunohistochemical detection of M1 (CD68, iNOS) and M2 (CD68, CD206) macrophage expression in arthritic tissues B ) Detection of TNF-α, IL-1β, IL-10 levels in arthritic tissues by ELISA. The descriptive statistics are presented as mean ±SD. * p < 0.05 vs. the CIA group, n = 3

Article Snippet: We measured the levels of serum markers using ELISA with a specialized commercial human IL-9 ELISA Kit (CSB-E04642h, Cusabio, Wuhan, China), human Gal1 ELISA Kit (CSB-EL012882h, Cusabio), human anti-CII ELISA Kit (HB3371-Hu, Shanghai Hengyuan Biotechnology Co., Ltd, Shanghai, China), human sSR-A ELISA Kit (HB3370-Hu, Shanghai Hengyuan Biotechnology Co., Ltd), human anti-Sa ELISA Kit (ml382511V), human RF ELISA Kit (ml060678V), human ACPA ELISA Kit (ml233305V), rat IL-9 ELISA Kit (CSB-E07294r, Cusabio), rat Gal1 ELISA Kit (JL25762, Shanghai Jianglai Biotechnology Co., Ltd, Shanghai, China), rat anti-CII ELISA Kit (HB1364-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat sSR-A ELISA Kit (HB1363-Ra, Shanghai Hengyuan Biotechnology Co., Ltd), rat anti-Sa ELISA Kit (ml202241V), rat RF ELISA Kit (ml059541V), and rat ACPA ELISA Kit (ml201458V).

Techniques: Inhibition, Activation Assay, Immunohistochemical staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: European journal of immunology

Article Title: Toll like Receptor 2 engagement on CD4 + T cells promotes TH9 differentiation and function

doi: 10.1002/eji.201646846

Figure Lengend Snippet: TLR2, but no other TLR ligands increase TGF-β and IL-4 driven IL-9 cytokine secretion. Naive CD4+ T cells were activated with anti-CD3 and anti-CD28 mAbs in the presence of different TLR ligands (1μg/ml) under non-polarizing and TH9 polarizing conditions for 48 h. IL-9 (A) and IFN-γ (B) ELISA were measured by culture supernatants. Means ± SD of three independent experiments are shown. * p < 0.05, ** p < 0.01, NS denotes non-significant.

Article Snippet: IFN-γ was measured by sandwich ELISA (R&D Systems; MIF00) and IL-9 ELISA was done using the DuoSet ELISA kit (R&D Systems; DY409) by following manufactured protocol.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: European journal of immunology

Article Title: Toll like Receptor 2 engagement on CD4 + T cells promotes TH9 differentiation and function

doi: 10.1002/eji.201646846

Figure Lengend Snippet: TLR2 engagement on Antigen85B specific CD4+ T cells increases TH9 differentiation driven by TGF-β and IL-4. Naive Ag85B transgenic CD4+ T cells were co-incubated with TLR2 KO BMDM pulsed with Ag85B peptide (1μg/ml) and co-stimulated with or without TLR2 ligand (P3CSK4) under non-polarizing and TH9 polarizing conditions for 48 h. (A) CD4+ T cells were permeabilized and labeled with mAbs to IL-9 and IFN-γ and percentages of IL-9+ or IFN-γ+ cells were determined by flow cytometry. (B, C) IL-9 and IFN-γ were measured in culture supernatants by ELISA. Means ± SD of five independent experiments are shown. * p < 0.05, ** p < 0.01. NS denote non- significant.

Article Snippet: IFN-γ was measured by sandwich ELISA (R&D Systems; MIF00) and IL-9 ELISA was done using the DuoSet ELISA kit (R&D Systems; DY409) by following manufactured protocol.

Techniques: Transgenic Assay, Incubation, Labeling, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: European journal of immunology

Article Title: Toll like Receptor 2 engagement on CD4 + T cells promotes TH9 differentiation and function

doi: 10.1002/eji.201646846

Figure Lengend Snippet: TLR2 engagement on human CD4+ T cells enhances IL9 mRNA and protein expression driven by polyclonal activation and TGF-β and IL-4. Naïve CD4+ T cells from three human donors were stimulated with anti-CD3 (10ug/ml) and anti-CD28 mAbs (1ug/ml) and exogenous TGF-β (5ng/ml) and IL-4 (10ng/ml), with or without P3CSK4 (2μg/ml) for 48h. (A) Relative expression of Il9 mRNA under non-polarizing and polarizing conditions with or without P3CSK4 (n=3) is shown. (B) IL-9 cytokine in culture supernatants were determined by ELISA. Means ± SD of three technical replicates for each donor are shown.

Article Snippet: IFN-γ was measured by sandwich ELISA (R&D Systems; MIF00) and IL-9 ELISA was done using the DuoSet ELISA kit (R&D Systems; DY409) by following manufactured protocol.

Techniques: Expressing, Activation Assay, Enzyme-linked Immunosorbent Assay