Human Chitinase 3-like 1/YKL-40 Antibody

Detection of Human Chitinase 3-like 1/YKL-40 by Western Blot. Western blot shows lysates of THP-1 human acute monocytic leukemia cell line treated (+) with 50 ng/mL of PMA for 72 hours. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Chitinase 3-like 1/YKL-40 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2599) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for Chitinase 3-like 1/YKL-40 at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Chitinase 3‑like 1/YKL-40 in Human Ovarian Cancer. Chitinase 3-like 1/YKL-40 was detected in immersion fixed paraffin-embedded sections of human ovarian cancer using Goat Anti-Human Chitinase 3-like 1/YKL-40 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2599) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Chitinase 3‑like 1/YKL-40 in THP‑1 Human Cell Line. Chitinase 3‑like 1/YKL-40 was detected in immersion fixed THP‑1 human acute monocytic leukemia cell line treated with PMA (positive staining) and THP‑1 human acute monocytic leukemia cell line (untreated; negative staining) using Goat Anti-Human Chitinase 3‑like 1/YKL-40 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2599) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Chitinase 3‑like 1/YKL-40 in Human Cartilage. Chitinase 3-like 1/YKL-40 was detected in immersion fixed paraffin-embedded sections of human cartilage using Goat Anti-Human Chitinase 3-like 1/YKL-40 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2599) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to chondrocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human Chitinase 3‑like 1/YKL-40 by Western Blot. Western blot shows lysates of human bone marrow, mouse bone marrow. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human Chitinase 3‑like 1/YKL-40 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2599) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Chitinase 3‑like 1/YKL-40 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of Human Chitinase 3‑like 1/YKL-40 by Simple Western^TM. Simple Western lane view shows lysates of Human bone marrow, loaded at 0.2 mg/mL. A specific band was detected for Chitinase 3‑like 1 at approximately 50 kDa (as indicated) using 25 µg/mL of Goat Anti-Human Chitinase 3‑like 1/YKL-40 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2599). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human Chitinase 3‑like 1/YKL-40 by Simple Western^TM. Simple Western lane view shows lysates of THP-1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 24 hours and 10 ug/ml LPS for 3 hours, loaded at 0.2 mg/mL. A specific band was detected for Chitinase 3‑like 1/YKL-40 at approximately 50 kDa (as indicated) using 25 µg/mL of Goat Anti-Human Chitinase 3‑like 1/YKL-40 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2599). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human Chitinase 3-like 1 by Western Blot YKL-40 expression in AD brain tissue (a) RT-qPCR analysis of YKL-40 in the frontal cortex of control, AD (I-III), AD (IV-VI) and rpAD (IV-VI) samples. GAPDH was used for normalization. Kruskal-Wallis and Dunn’s post-hoc tests were used to estimate statistical differences. b Western blot analysis of YKL-40 in the frontal cortex of control, AD (IV-VI) and rpAD (IV-VI) samples. Normalization was based on GAPDH levels. Graphic summary of densitometry analyses performed on western blot results acquired from 8 control, 8 AD and 6 rpAD samples. c Immunohistochemical analysis of YKL-40 in the cerebral cortex, white matter, subpial layer and cerebellum in control and AD cases. d Immunohistochemical analysis of YKL-40 in the temporal cortex and hippocampus in AD cases. e Immunohistochemical analysis of YKL-40+ astrocytes surrounding beta -amyloid plaques (left) and in blood vessels with amyloid angiopathy (right) in the hippocampal region of AD cases Brown staining corresponds to YKL-40 staining and light blue to haematoxylin counterstaining. f Double-labeling immunofluorescence of YKL-40 (green) and amyloid beta (red) in the hippocampus of AD. g Double-labeling immunofluorescence of YKL-40 (green) and GFAP (red) in cerebral cortex and white matter in AD tissues. Fold changes in expression of mRNA and protein were determined relative to the control cases. *p < 0.05, ***p < 0.001 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29126445), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Chitinase 3-like 1 by Western Blot YKL-40 expression in DLB brain tissue. a RT-qPCR analysis of YKL-40 in the frontal cortex of control, DLB and rpDLB samples. Normalization was based on GAPDH levels. b Western blot analysis of YKL-40 in the frontal cortex of control, DLB and rpDLB samples. Normalization was carried out with GAPDH. Kruskal-Wallis and Dunn’s post-hoc tests were used for determination of statistical differences. c Immunohistochemical analysis of YKL-40 in the cerebral cortex, white matter and subpial layer in DLB cases. Brown staining corresponds to YKL-40 staining and light blue to haematoxylin counterstaining. Fold changes in expression (mRNA and protein) were determined relative to the control cases Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29126445), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Chitinase 3-like 1 by Western Blot YKL-40 expression in the brain tissue of sCJD and related mouse model. a RT-qPCR analysis of YKL-40 in the frontal cortex (left panel) and cerebellum (right panel) of control, sCJD MM1 and sCJD VV2 samples. GAPDH was used for normalization. b Western blot analysis of YKL-40 in the frontal cortex (upper panel) and cerebellum (bottom panel) of control, sCJD MM1 and sCJD VV2 samples. For normalization GAPDH was used. Graphical representation of Western Blot data acquired from the analysis of eight samples per group. Fold changes in the expression of mRNA and protein were determined relative to the control cases. Kruskal-Wallis and Dunn’s post-hoc tests were used to determine statistical differences. ***p < 0.001 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29126445), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Chitinase 3-like 1 by Western Blot YKL-40 expression in experimental models of prion diseases. a RT-qPCR analysis of YKL-40 in the cortex of control and sCJD MM1 inoculated tg340PRNP129MM mice at 120 dpi (pre-clinical), 160 dpi (early clinical), 180 dpi (clinical) and 210 dpi (clinical with 10–1 diluted inoculum). Four animals per group were analyzed. Normalization was performed using Hprt. b Representative Western-blot analyses for YKL-40 immunodetection in the cortex of control and sCJD MM1-inoculated tg340PRNP129MM mice at 120 dpi (pre-clinical) and 180 dpi (clinical). Three animals per group were analyzed. Normalization was based on beta -actin levels. Numbers indicate densitometry results from three animals per group. Unpaired t-tests were performed to determine statistical differences. c RT-qPCR analysis of YKL-40 in the whole brain of control and RML scrapie-infected mice at pre-clinical (120 dpi) and clinical disease (180 dpi) stages. GAPDH was used for normalization. Similar results were acquired when normalization was based on Hprt expression levels (not shown). Unpaired t-tests were used for estimation of statistical differences. d Immunohistochemical analysis of YKL-40 expression in the cerebral cortex, hippocampus and thalamus of control and RML scrapie-infected mice at pre-clinical (60 and 90 dpi) and clinical (150 dpi) disease stages. Scale bar = 50 μm. Arrows indicate YKL-40 positive reactive astrocytes. Three animals per time point were used. Two sections were stained per animal (sagittal and coronal sections were used). Brown staining corresponds to YKL-40 staining and light blue to haematoxylin counterstaining. e RT-qPCR analysis of YKL-40 in the cerebral cortex of control and 22 L scrapie-infected mice at pre-clinical (60 dpi) and clinical (140 dpi) disease stages. f RT-qPCR analysis of YKL-40 in the cerebellum of control and 22 L scrapie-infected mice at clinical (140 dpi) disease stages. In all cases GAPDH was used for normalization. Similar results were obtained when Hprt was used for normalization (not shown). Unpaired t-tests were used to determine statistical differences. g Western blot analysis of Vimentin and GFAP in control and 22 L–scrapie infected COCS. beta -actin was used for normalization. Graphic representation of densitometry analysis of five cerebellar tissues per condition is shown. Statistical differences were determined with unpaired t-tests. h RT-QuIC analysis of control and 22 L–scrapie infected COCS. A representative graph of three cerebellar tissues per condition is shown. All control tissues were tested negative, while all 22 L–infected tissues were tested positive (left panel); western-blot showing the presence of PrPres in 22 L–infected cells (right panel). i ELISA analysis of YKL-40 levels in control and 22 L–scrapie infected COCS. Statistical differences were determined with unpaired t-tests. Fold changes in the expression of mRNA and protein were determined relative to the control cases. *p < 0.05, **p < 0.01, ***p < 0.001 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29126445), licensed under a CC-BY license. Not internally tested by R&D Systems.