Chitinase 3-like 1/YKL-40 Antibody [Unconjugated]

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 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 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 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.

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.

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.

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.

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 (//pubmed.ncbi.nlm.nih.gov/29126445), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/29126445), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/29126445), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/29126445), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Chitinase 3-like 1/YKL-40 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Chitinase 3-like 1/YKL-40
  Tyr22-Thr383
  Accession # P36222
• Specificity: Detects human Chitinase 3-like 1/YKL-40 in direct ELISAs and Western blots. In direct ELISAs, approximately 15% cross‑reactivity with recombinant mouse Chitinase 3-like 1/YKL-40 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CHI3L1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 1.5-15 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Simple Western 10-25 ug/mL
  • Western Blot 0.5-1.0 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Chitinase 3-like 1/YKL-40 Antibody [Unconjugated]

• AW208766
• Brp39
• CHI3L1
• Chitinase 3 like 1
• Chitinase 3-like 1
• gp39
• HCgp39
• YKL-40

Background

CHI3L1, also known as HCgp39 (human cartilage glycoprotein 39) and YKL40, is a secreted glycoprotein belonging to the family of chitinase-like lectins. These proteins are structurally related to the glycosylhydrolase family 18, but lack enzymatic activity. CHI3L1 is expressed by chondrocytes, synovial cells, macrophages and neutrophils. It has been found to play a role in down‑regulating cytokine-induced inflammatory responses. Human CHI3L1 shares 73%, 80%, 83% and 86% amino acid sequence identity with mouse, rat, bovine and porcine CHI3L1, respectively.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Chitinase 3-like 1/YKL-40 Antibody (AF2599)(23)

Publications using AF2599

• Jiang, W;Zhu, F;Xu, H;Xu, L;Li, H;Yang, X;Khan Afridi, S;Lai, S;Qiu, X;Liu, C;Li, H;Long, Y;Wang, Y;Connolly, K;Elias, JA;Lee, CG;Cui, Y;Huang, YA;Qiu, W;Tang, C; CHI3L1 signaling impairs hippocampal neurogenesis and cognitive function in autoimmune-mediated neuroinflammation Science advances 2023-09-29 [PMID: 37756391] (Immunohistochemistry, Mouse)
• Li J, Jaiswal MK, Chien JF et al. Divergent single cell transcriptome and epigenome alterations in ALS and FTD patients with C9orf72 mutation Nat Commun 2023-09-15 [PMID: 37714849] (Simple Western, Human)
• Bárbara Alves, Joana Peixoto, Sofia Macedo, Jorge Pinheiro, Bruno Carvalho, Paula Soares et al. High VEGFA Expression Is Associated with Improved Progression-Free Survival after Bevacizumab Treatment in Recurrent Glioblastoma Cancers (Basel) 4/7/2023 [PMID: 37190125]
• Bárbara Alves, Joana Peixoto, Sofia Macedo, Jorge Pinheiro, Bruno Carvalho, Paula Soares et al. High VEGFA Expression Is Associated with Improved Progression-Free Survival after Bevacizumab Treatment in Recurrent Glioblastoma Cancers (Basel) 2023-04-07 [PMID: 37190125] (Immunohistochemistry, Human)
• Wei Wei, Yuezhang Wu, Dong-Dong Chen, Yuntao Song, Guohui Xu, Qi Shi et al. Proteomics profiling for the global and acetylated proteins of papillary thyroid cancers Proteome Science 4/26/2023 [PMID: 37101287]
• Wenjin Qiu, Zumu Xiao, Yushi Yang, Lishi Jiang, Shibin Song, Xiaolan Qi et al. USP10 deubiquitinates RUNX1 and promotes proneural-to-mesenchymal transition in glioblastoma Cell Death & Disease 3/22/2023 [PMID: 36949071]
• A Huss, A Abdelhak, B Mayer, H Tumani, HP Müller, K Althaus, J Kassubek, M Otto, AC Ludolph, D Yilmazer-H, H Neugebauer Association of Serum GFAP with Functional and Neurocognitive Outcome in Sporadic Small Vessel Disease Biomedicines, 2022-08-02;10(8):. 2022-08-02 [PMID: 36009416] (IHC, Human)
• Yanaika S. Hok-A-Hin, Jeroen J. M. Hoozemans, William T. Hu, Dorine Wouters, Jennifer C. Howell, Alberto Rábano et al. YKL-40 changes are not detected in post-mortem brain of patients with Alzheimer’s disease and frontotemporal lobar degeneration Alzheimer's Research & Therapy 12/1/2022 [PMID: 35879733]
• Laura Cubas-Núñez, Sara Gil-Perotín, Jéssica Castillo-Villalba, Verónica López, Luis Solís Tarazona, Raquel Gasqué-Rubio et al. Potential Role of CHI3L1+ Astrocytes in Progression in MS Neurology - Neuroimmunology Neuroinflammation 5/4/2021 [PMID: 33658322]
• Siqi Xiong, Ajay Kumar, Shenghe Tian, Eman E Taher, Enzhi Yang, Paul R Kinchington et al. Stem cell transplantation rescued a primary open-angle glaucoma mouse model eLife 1/28/2021 [PMID: 33506763]
• S Zhang, A Sousa, M Lin, A Iwano, R Jain, B Ma, CM Lee, JW Park, S Kamle, R Carlson, GG Lee, JA Elias, JR Wands Role of Chitinase 3-Like 1 Protein in the Pathogenesis of Hepatic Insulin Resistance in Nonalcoholic Fatty Liver Disease Cells, 2021-01-20;10(2):. 2021-01-20 [PMID: 33498326] (IHC, Human)
• JOANNA SALOMON, ALEKSANDRA PIOTROWSKA, ŁUKASZ MATUSIAK, PIOTR DZIĘGIEL, JACEK C. SZEPIETOWSKI Chitinase-3-like Protein 1 (YKL-40) Is Expressed in Lesional Skin in Hidradenitis Suppurativa In Vivo 2019-01-01 [PMID: 30587614] (Immunohistochemistry, Human)
• JOANNA SALOMON, ALEKSANDRA PIOTROWSKA, ŁUKASZ MATUSIAK, PIOTR DZIĘGIEL, JACEK C. SZEPIETOWSKI Chitinase-3-like Protein 1 (YKL-40) Is Expressed in Lesional Skin in Hidradenitis Suppurativa In Vivo 1/1/2019 [PMID: 30587614]
• JOANNA SALOMON, ALEKSANDRA PIOTROWSKA, ŁUKASZ MATUSIAK, PIOTR DZIĘGIEL, JACEK C. SZEPIETOWSKI Chitinase-3-like Protein 1 (YKL-40) Expression in Squamous Cell Skin Cancer Anticancer Research 8/1/2018 [PMID: 30061245]
• M Querol-Vil, M Colom-Cade, J Pegueroles, C San Martín, J Clarimon, O Belbin, J Fortea, A Lleó YKL-40 (Chitinase 3-like I) is expressed in a subset of astrocytes in Alzheimer&#039;s disease and other tauopathies J Neuroinflammation, 2017-06-09;14(1):118. 2017-06-09 [PMID: 28599675] (IHC, Human)
• SR Langley, K Willeit, A Didangelos, LP Matic, P Skroblin, J Barallobre, M Lengquist, G Rungger, A Kapustin, L Kedenko, C Molenaar, R Lu, T Barwari, G Suna, X Yin, B Iglseder, B Paulweber, P Willeit, J Shalhoub, G Pasterkamp, AH Davies, C Monaco, U Hedin, CM Shanahan, J Willeit, S Kiechl, M Mayr Extracellular matrix proteomics identifies molecular signature of symptomatic carotid plaques J. Clin. Invest, 2017-03-20;0(0):. 2017-03-20 [PMID: 28319050] (IHC, Human)
• Katherine Xu, Paul Rosenstiel, Neal Paragas, Christian Hinze, Xiaobo Gao, Tian Huai Shen et al. Unique Transcriptional Programs Identify Subtypes of AKI Journal of the American Society of Nephrology 6/1/2017 [PMID: 28028135]
• Serum YKL-40 as a marker of liver fibrosis in patients with non-alcoholic fatty liver disease Sci Rep, 2016-10-14;6(0):35282. 2016-10-14 [PMID: 27739482] (IHC, Human)
• Kunz L, Van't Wout E, van Schadewijk A, Postma D, Kerstjens H, Sterk P, Hiemstra P Regulation of YKL-40 expression by corticosteroids: effect on pro-inflammatory macrophages in vitro and its modulation in COPD in vivo. Respir Res, 2015-12-22;16(0):154. 2015-12-22 [PMID: 26696093] (Flow Cytometry, IHC-Fr, Human)
• Du Y, Yun H, Yang E, Schuman J Stem cells from trabecular meshwork home to TM tissue in vivo. Invest Ophthalmol Vis Sci, 2013-02-19;54(2):1450-9. 2013-02-19 [PMID: 23341019] (IHC, Human)
• Catherine A. Formolo, Russell Williams, Heather Gordish-Dressman, Tobey J. MacDonald, Norman H. Lee, Yetrib Hathout Secretome Signature of Invasive Glioblastoma Multiforme Journal of Proteome Research 7/1/2011 [PMID: 21574646]
• Park JA, Drazen JM, Tschumperlin DJ The chitinase-like protein YKL-40 is secreted by airway epithelial cells at base line and in response to compressive mechanical stress. J. Biol. Chem., 2010-07-22;285(39):29817-25. 2010-07-22 [PMID: 20650887] (Western Blot, Human)
• Agapov E, Battaile JT, Tidwell R, Hachem R, Patterson GA, Pierce RA, Atkinson JJ, Holtzman MJ Macrophage chitinase 1 stratifies chronic obstructive lung disease. Am. J. Respir. Cell Mol. Biol., 2009-06-02;41(4):379-84. 2009-06-02 [PMID: 19491341] (IHC-P, Human)

Bioinformatics

• Gene Symbol: CHI3L1
• Uniprot:
  • P36222()