human saa1 elisa kit  (R&D Systems)

Journal: Nature Communications

Article Title: CD8 + T cell loss induces autoinflammation in inborn errors of cell death

doi: 10.1038/s41467-026-70317-2

Figure Lengend Snippet: a , b Schematic diagram showing serum cytokine microarray analysis of continuous P2’s serum samples before (PS1 and PS2) or after (PS3, flare; PS4 and PS5, non-flare) the onset of recurrent fever ( a ). PCA analysis of sample variance determined by semi-quantitative 174 cytokines antibody array on serum samples of C1-C3 from three unaffected controls and PS1-PS5 from P2 ( b ). c–e The integrated relative MFI plots of P2’s serum cytokine levels. 26 cytokines peaked in PS3 were enrich in group 1 (left), 24 cytokines higher than control were enriched in group 2 (right) ( c ). The relative MFI was calculated by normalizing the average MFI of the unaffected controls ( n = 3) to 0, and setting the largest MFI to 1 or the smallest MFI to −1 across 5 samples from P2. Boxplots exhibiting expression scores of cytokines of group 1 ( d ) and group 2 ( e ) in indicated cell with Seurat AddModuleScore function. The cell number of each subtype (listed in the “Methods” section), was employed for p value calculations. Boxes show the median (center line), 25th and 75th percentiles (upper and lower hinges), and whiskers extend to 1.5× the interquartile range, representing the maximum and minimum values. f Average expression values (arcsinh transformed) of cl. Casp-3 across identified peripheral immune cell lineages in PBMCs freshly isolated from four unaffected controls (C) and two patients (P, 1 sample from P1 and 2 samples from P2) by mass cytometry by time-of-flight (CyTOF) analysis. g–j Representative contour plots of cl. Casp-3 ( g , h ) and histograms of total RIPK1, pRIPK1(S166) levels ( i , j ) in CD4 + T and CD8 + T cells in freshly isolated PBMCs from unaffected controls (for cl. Casp-3, n = 12; for pRIPK1(S166), n = 10) and patients ( n = 4 for P1 and P2) (left). Summary histograms of cl. Casp-3 and pRIPK1(S166) (right) were shown. k Representative histogram and statistical analysis of surface CD69 expression on CD3 + T cells from unaffected controls ( n = 7) and two patients ( n = 3 for P1 and P2). l Serum IFNγ, sCD25, sCD95 and CD95L of unaffected controls (for IFNγ, n = 9; for sCD25, n = 15; for sCD95 and CD95L, n = 10), two patients (for IFNγ, P1, n = 5, P2 = 6; for sCD25, P1, n = 5, P2, n = 4; for sCD95, P1 and P2, n = 7; for CD95L, P1 = 7; P2 = 8) and their parents (for IFNγ and sCD25, n = 7; for sCD95 and CD95L, n = 6) as well as two CRIA syndrome patients ( n = 6) determined by ELISA. T denotes 10 ng ml -1 TNF; S denotes 250 nM SM164; Z denotes 25 μM z-VAD-fmk. All histogram graphs show mean ± SD. P values were determined by two-tailed unpaired t-test. Data in g–k were pooled from two independent experiments.

Article Snippet: Human or mouse serum cytokines were measured by enzyme-linked immunosorbent assay (ELISA) (Multi Sciences: 70-EK1280, 70-EK194, 70-EK182HS, 70-EK106HS, 70-EK191, 70-EK180HS, 70-EK1F01, 70-EK1F02, EK206HS, EK282HS/2) according to the manufacturer’s instructions.

Techniques: Microarray, Ab Array, Control, Expressing, Transformation Assay, Isolation, Mass Cytometry, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Nature Communications

Article Title: CD8 + T cell loss induces autoinflammation in inborn errors of cell death

doi: 10.1038/s41467-026-70317-2

Figure Lengend Snippet: a Schematic diagram of T cell-MDM co-culture system. b , c qRT-PCR analysis of TNF, IL1B and IL6 mRNA levels in GM-CSF-activated monocytes derived macrophages (MDMs) co-cultured with or without T cells for 6 h. GM-CSF-activated MDMs generated from ( b , left) unaffected control 1 (C1) or ( b , right) unaffected control 3 (C3) or P1 ( c ) were co-cultured with donor-matched T cells as well as T cells from other unaffected controls (C2 and C4) or patients as indicated. d Dot plot of IFNG, TNF in cell subtypes found in scRNA-seq of PBMCs from three unaffected controls and two patients. e Representative histograms and statistical analysis of TNF and IFNγ expression in CD8 + T cells in PBMCs isolated from unaffected controls ( n = 6) and patients ( n = 3 from P1 and P2) treated with 10 ng ml -1 TNF (T) with or without 250 nM SM-164 (S), 25 μM z-VAD-fmk (Z) for 24 h. f , T cells isolated from P2 were co-cultured with MDMs generated from unaffected control in the presence of 10 μg ml -1 adalimumab, 10 μg ml -1 emapalumab alone or in combination for 6 h. TNF, IL1B and IL6 mRNA levels of MDMs were analyzed by qRT-PCR. g Experimental timeline for generation of lineage negative bone marrow cells (Lin - BMs)-transplanted mice with conditional overexpression of human WT or mutated RIPK1 K377E or R390G in CD8 + T cell- or myeloid cell-specific lineage. h, i ELISA measurement of serum Tnf and Il6 levels in mice specifically overexpressing WT or mutated hRIPK1 in ( h ) CD8 + T cells (EV, n = 5; WT, K377E and R390G n = 7) or ( i ) myeloid cells (EV, n = 5; WT, K377E and R390G n = 6). All histogram graphs show mean ± SD. P values were determined by one-way ANOVA with Tukey’s multiple comparisons test. Data in b , c , and f show the means ± SD of technical triplicates. The results shown in b were derived from two independent experiments using cells from different donors. The experiment in ( c , h , i , f ) is representative of two independent experiments. Data in e were pooled from two independent experiments.

Article Snippet: Human or mouse serum cytokines were measured by enzyme-linked immunosorbent assay (ELISA) (Multi Sciences: 70-EK1280, 70-EK194, 70-EK182HS, 70-EK106HS, 70-EK191, 70-EK180HS, 70-EK1F01, 70-EK1F02, EK206HS, EK282HS/2) according to the manufacturer’s instructions.

Techniques: Co-Culture Assay, Quantitative RT-PCR, Derivative Assay, Cell Culture, Generated, Control, Expressing, Isolation, Over Expression, Enzyme-linked Immunosorbent Assay

Journal: Journal of Neuroinflammation

Article Title: Single-cell RNA sequencing uncovers neutrophil clusters associated with autoimmune neuroinflammation

doi: 10.1186/s12974-026-03772-9

Figure Lengend Snippet: Enhanced SAA1 expression and NET formation in plasma of MS patients. (A) SAA1 protein expression was quantified by ELISA in plasma samples collected from MS patients and HC. (B) H3.1-nucleosome expression was quantified by ELISA in plasma samples collected from MS patients and HC

Article Snippet: The human SAA1 ELISA kit (DY3019-05; R & D Systems, Minneapolis, MN) detected human SAA1 in plasma samples from MS patients and HC.

Techniques: Expressing, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Journal of Advanced Research

Article Title: Characterization of immune features and discovery of potential biomarkers for ankylosing spondylitis using deep plasma proteomics

doi: 10.1016/j.jare.2025.05.052

Figure Lengend Snippet: Verification of potential biomarkers for AS. (A) Expression levels of SAA1, FERMT3, ILK, and TLN1 in plasma samples from active-stage AS patients, stable-stage AS patients, and healthy controls measured by proteomics. ROC curves for the individual protein (SAA1, FERMT3, ILK, and TLN1) as a predictor to classify AS patients from healthy controls (B), and active-stage AS patients from stable-stage AS patients (C). (D) ELISA analysis of SAA1, FERMT3, ILK, and TLN1 expressions in the plasma samples from an independent validation cohort of healthy controls (N, n = 24), active-stage AS (A, n = 27), and stable-stage AS patients (S, n = 28). Statistical analyses employed Wilcoxon rank-sum test for intergroup comparisons, with results reported as mean ± SEM. Significance thresholds based on BH-adjusted p -values were defined as: *, <0.05; **, <0.01; ***, <0.001.

Article Snippet: The human serum amyloid A (SAA1) ELISA kit (#OKIA00083) was purchased from Aviva Systems Biology Company (San Diego, CA, USA).

Techniques: Expressing, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Biomarker Discovery

Journal: Arthritis Research & Therapy

Article Title: Proteomic profiling identifies SPP1 associated with rapidly progressive interstitial lung disease in anti-MDA5-positive dermatomyositis

doi: 10.1186/s13075-023-03243-z

Figure Lengend Snippet: Plasma concentration of SPP1, SAA1, and KNG1 in MDA5 + DM RP-ILD. A–C The concentrations of SPP1, SAA1, and KNG1 in the plasma from an independent cohort were determined by ELISA. P values were calculated by the ANOVA test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. D–F ROC curve showed the AUC of the 3 verified proteins, respectively

Article Snippet: Following the manufacturer’s instructions, commercial ELISA kits were utilized for quantifying the levels of serum amyloid A1 (SAA1) (Boster Bio, China), V-Set And Immunoglobulin Domain Containing 4 (VSIG4), Coagulation Factor XI (F11) (both from CUSABIO, China), Coagulation Factor XIII A Chain (F13A1), Secreted phosphoprotein 1 (SPP1) and Kininogen 1 (KNG1) (all from Uscn life Science, China) in plasma samples from both patients and normal control subjects.

Techniques: Clinical Proteomics, Concentration Assay, Enzyme-linked Immunosorbent Assay