human saa1 elisa kit  (R&D Systems)

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: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: TGFβ1 signaling in primary CRC and LM-CRC CAFs induces expression of IL-6 (A) Schematic overview of workflow for CAF isolation and TGFβ1 pathway-targeted qPCR array. (B) RNA expression of TGFβ1 target genes in two primary CRC CAFs (CAF1 and CAF2) and one LM-CRC CAF. qPCR values have been log transformed. (C) Numerical values depicting the seven TGFβ1 target genes that showed consistent upregulation. (D) Fold change RNA expression of these seven TGFβ1 target genes (yellow symbols). (E) IL6 RNA expression in primary CRC CAFs (CAF4 and CAF5) and the CCD-18Co colon fibroblast line after stimulation with TGFβ1 or medium control. N = 3 CCD-18Co, CAF4; N = 2 CAF5 independent biological experiments. ∗∗ p ≤ 0.01 determined by paired T test. (F) Protein expression of IL-6 in tissue lysates of normal colon and primary CRC (left panel, N = 50) or normal liver and LM-CRC (right panel, N = 50) as determined by ELISA. ∗∗∗∗ p ≤ 0.0001, ∗ p ≤ 0.05 determined by unpaired T test. (G) IL6 RNA expression in unstimulated normal colon fibroblasts ( N = 8) or primary CRC CAFs ( N = 10). (H) RNA expression of IL6 in the original CMS classified cohort (CMS1: N = 457; CMS2: N = 1183; CMS3: N = 409; CMS4: N = 773). ∗∗∗∗ p ≤ 0.0001 determined by one-way ANOVA with correction for multiple testing (Dunnett’s test).

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: Expressing, Isolation, RNA Expression, Transformation Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: IL-6 signaling in CAFs is partly regulated through autocrine TGFβ1 signaling (A) RNA expression of IL6 in CAFs derived either from the primary tumor (CRC CAF, N = 18) or liver metastasis (LM-CRC CAF, N = 6). ∗∗∗ p ≤ 0.001 determined by unpaired T test. (B) RNA-seq data from GSE46824 dataset showing IL6 RNA expression in an independent cohort of primary CRC CAFs ( N = 14) and LM-CRC CAFs ( N = 11). ∗∗∗ p ≤ 0.001 determined by unpaired T test. (C) IL-6 protein expression in unstimulated primary CRC CAF ( N = 8) and LM-CRC CAF ( N = 6) supernatant determined by ELISA. ∗∗ p ≤ 0.01 determined by unpaired T test. (D and E) IL-6 protein expression in TGFβ1-stimulated (5 ng/mL) primary CRC CAFs ( N = 8) (D) and LM-CRC CAFs ( N = 6) (E). ∗ p ≤ 0.05 determined by unpaired T test. (F) Fold change IL-6 protein expression in supernatant from TGFβ1-stimulated vs. unstimulated primary CRC CAFs ( N = 8) and LM-CRC CAFs ( N = 6). ns, p > 0.05 determined by unpaired T test. (G) Percentage decrease in IL-6 protein expression in supernatant in primary CRC ( N = 5) and LM-CRC ( N = 5) CAFs after inhibition with SB431532 (Alk5 inhibitor). ns, p > 0.05 determined by unpaired T test. (H) RNA-seq data from GSE46824 dataset (2B) showing TGFβ1 RNA expression in an independent cohort of primary CRC CAFs ( N = 14) and LM-CRC CAFs ( N = 11). ∗ p ≤ 0.05 determined by unpaired T test. (I) TGFβ1 protein expression in unstimulated primary CRC CAF ( N = 5) and LM-CRC CAF ( N = 6) supernatant determined by ELISA. ∗ p ≤ 0.05 determined by unpaired T test.

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: RNA Expression, Derivative Assay, RNA Sequencing, Expressing, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: TGFβ isoforms are elevated in CMS4 subtype CRC and are capable of inducing IL-6 expression (A–C) RNA expression of the three TGFβ isoforms in the original CMS cohort (CMS1: N = 457; CMS2: N = 1183; CMS3: N = 409; CMS4: N = 773). ∗∗∗∗ p ≤ 0.0001 determined by one-way ANOVA with correction for multiple testing (Dunnett’s test). (D) Protein expression of TGFβ isoforms in the supernatant of primary CRC and LM-CRC CAFs 48 h after the start of serum starvation as determined by ELISA. (E and F) Expression of IL-6 in supernatant of primary CRC CAFs ( N = 7) (E) or LM-CRC CAFs ( N = 6) (F) 48 h after start stimulation with either TGFβ2 or TGFβ3. ∗ p ≤ 0.05 determined by paired T test.

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: Expressing, RNA Expression, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: TGFβ-mediated IL-6 production by CAFs induces the expression of neutrophil chemoattractants SAA1 and CXCL5 in hepatocytes (A) Schematic overview of ‘CAF priming of Huh-7 hepatocytes’, referring to Huh-7 cells that are exposed to TGFβ1-stimulated CRC CAF CM. (B) Western blot for pSTAT3 and total STAT3 in wildtype Huh-7 cells that were stimulated with CAF CM or TGFβ1 CAF CM from 3 different CRC CAF lines. (C) RNA expression of SAA1 and CXCL5 in wildtype Huh-7 cells stimulated with DMEM 0% FCS (Control), IL-6 (50 ng/mL), CAF CM, or TGFβ1 CAF CM. N = 2 or N = 3 for the Control group, due to the very low SAA1 expression; for the other conditions, N = 3.∗∗∗ p ≤ 0.001, ∗∗ p ≤ 0.01, ∗ p ≤ 0.05 determined by one-way ANOVA with correction for multiple testing (Dunnett’s test). (D and E) RNA expression of SAA1 and CXCL5 in wildtype Huh-7 cells after CAF priming with blockade of specific components of the IL-6 signaling pathway. Siltuximab (anti-IL-6, 2 μg/mL), tocilizumab (anti-IL6R, 8 μg/mL), α-gp130 (anti-gp130, 8 μg/mL), tofacitinib (anti-JAK1/3, 5 μM), or human IgG isotype control (Bio X Cell; 2 μg/mL) were added to Huh-7 cells along with TGFβ1-stimulated CAF CM for 15 min. Subsequently, these different stimulations were applied to Huh-7 cells for 10 min. N = 1 for all different conditions. ∗∗∗∗ p ≤ 0.0001, ∗∗∗ p ≤ 0.001 determined by one-way ANOVA with correction for multiple testing (Dunnett’s test).

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: Expressing, Western Blot, RNA Expression, Control

Journal: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: The induction of neutrophil chemoattractants in hepatocytes is fully gp130-dependent (A) Western blot for pSTAT3 and total STAT3 in CAF CM-primed Huh-7 cells in which gp-130 signaling is blocked using α-gp130 antibody. α-gp130, anti-gp130. β-actin is used as a loading control. (B) Western blot for pSTAT3 and total STAT3 in vector control, gp130 KO , or gp130 KO rescued Huh-7 cells (gp130 KO+Rescue ) after stimulation with DMEM 0% (Control) or TGFβ1 CAF CM. (C) RNA expression of SAA1 in gp130 KO and gp130 KO+Rescue Huh-7 cells after CAF priming ( N = 3 independent biological experiments). ∗∗ p ≤ 0.01 determined by unpaired T test. (D) IHC of pSTAT3 in Huh-7 cells harboring a vector control, gp130 KO , or gp130 KO+Rescue after stimulation with DMEM 0% or after CAF priming. Scale bar, 50 μm.

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: Western Blot, Control, Plasmid Preparation, RNA Expression

Journal: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: The IL-6 family of cytokine-JAK-STAT signaling axis is active in CMS4 CRC in vivo (A) Schematic overview of the KPN GEMM organoid experiment. (B) Gene Set Enrichment analysis shows the Enrichment Score (ES) for the IL6-JAK-STAT signaling hallmark in KPN tumor tissue ( N = 3) and wildtype normal colon ( N = 3). (C) Normalized counts of IL6, IL11, and LIF in KPN tumor tissue (KPN) and normal colon tissue (WT). ∗∗∗∗ p ≤ 0.0001, ∗∗ p ≤ 0.01 determined by unpaired T test. (D) Quantification of automated scoring of Ly6G staining in pre-metastatic livers of KPN transplanted mice ( N = 4) and WT livers ( N = 5). (E) Representative IHC of pSTAT3, Ly6G, and GFP in pre-metastatic and metastatic livers in the KPN GEMM. Scale bar, 50 μm. (F) Waterfall plot of the difference in serum SAA1 OD450 value before and after surgery in 16 patients with primary CRC without liver metastasis.

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: In Vivo, Staining

Journal: BMJ Neurology Open

Article Title: Altered amyloid plasma profile in patients with disabling headaches after SARS-CoV-2 infection and vaccination

doi: 10.1136/bmjno-2024-001013

Figure Lengend Snippet: Plasma levels of (A) amyloid precursor protein (APP), (B) cathepsin L, (C) pregnancy zone protein (PZP) and (D) serum amyloid A (SAA1) in healthy controls (HC) (n=16), participants with persistent headache after SARS-CoV-2 vaccine (COvax) (n=31) and participants with persistent headache after COVID-19 ( C19 ) (n=29).

Article Snippet: Plasma levels of APP (Cat# DY850), PZP (Cat# DY8280-05), CTSL (Cat# DY952) and SAA1 (Cat# DY3019-05) were measured by ELISA using commercially available antibodies (R&D Systems, Minneapolis, Minnesota, USA) in a 384-format using a combination of a SELMA pipetting robot (Analytik Jena AG, Jena, Germany) and a BioTek dispenser/washer (BioTek Instruments, Winooski, VT).

Techniques: Clinical Proteomics

Journal: International journal of molecular sciences

Article Title: Elevated Serum Amyloid A Levels Contribute to Increased Platelet Adhesion in COVID-19 Patients.

doi: 10.3390/ijms232214243

Figure Lengend Snippet: Figure 1. SAA is bound to platelets from COVID-19 patients. (A) Mass spectrometry was used to identify and quantify proteins overexpressed in purified COVID-19 platelets compared to controls. Shown are proteins whose levels were consistently elevated in two independent experiments. Values indicate the average fold-change (FC) in the relative label-free quantification intensity between the COVID-19 and the control samples (n = 4 in each group). Unpaired Student’s t-test was used to determine the significance. (B) Platelet lysates from control or COVID-19 donors were separated using SDS-PAGE. SAA was detected using a specific antibody. Tubulin was used as a loading control. Results are representative of six controls and six COVID-19 patients. Numbers represent fold intensity of control densitometric values normalized to tubulin. (C) SAA levels in platelet lysates from controls (n = 17) or COVID-19 patients (n = 15) were determined using ELISA. The dashed line in the violin plot is the median. The dotted lines show the interquartile range. Unpaired t-test, **** p < 0.001. (D) Platelet lysates from control or COVID-19 donors separated according to disease severity were analyzed using SDS-PAGE (n = 6 controls, 9 mild, 5 moderate, and 20 severe disease patients). SAA was detected using a specific antibody. The results were normalized to tubulin and to the amount of total protein. Ordinary one-way ANOVA, Dunnett’s multiple comparisons compared to control, ns = non-significant, * p < 0.05.

Article Snippet: An SAA-specific ELISA (SAA1 DuoSet ELISA Kit, R&D Systems, MN USA) was used to detect and quantify the SAA levels in the platelets of healthy and COVID-19 patients according to the manufacturer’s instructions.

Techniques: Mass Spectrometry, Control, SDS Page, Enzyme-linked Immunosorbent Assay