重组Anti-YKL-40/CHI3L1抗体[EPR19078-157] (ab255297)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19078-157] to YKL-40/CHI3L1
- Suitable for: WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-YKL-40/CHI3L1抗体[EPR19078-157]
参阅全部 YKL-40/CHI3L1 一抗 -
描述
兔单克隆抗体[EPR19078-157] to YKL-40/CHI3L1 -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, Flow Cyt (Intra)more details
不适用于: ICC/IF -
种属反应性
与反应: Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Human spleen tissue lysate; THP-1 (Human monocytic leukemia monocyte), U-87 MG ( Human glioblastoma-astrocytoma epithelial cell), MG-63 (Human osteosarcoma fibroblast), Caco-2 (Human colorectal adenocarcinoma epithelial cell), 293T (Human embryonic kidney epithelial cell), and Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates; IHC-P: Human spleen and tonsil tissue. Flow: THP-1 cells.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR19078-157 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab255297于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB |
1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 43 kDa).
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IHC-P | (1) |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/500.
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说明 |
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WB
1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 43 kDa). |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/500. |
靶标
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功能
Carbohydrate-binding lectin with a preference for chitin. May play a role in defense against pathogens, or in tissue remodeling. May play an important role in the capacity of cells to respond to and cope with changes in their environment. -
组织特异性
Present in activated macrophages, articular chondrocytes, synovial cells as well as in liver. Undetectable in muscle tissues, lung, pancreas, mononuclear cells, or fibroblasts. -
疾病相关
A genetic variation in CHI3L1 is associated with susceptibility to asthma-related traits type 7 (ASRT7) [MIM:611960]. Asthma-related traits include clinical symptoms of asthma, such as coughing, wheezing and dyspnea, bronchial hyperresponsiveness (BHR) as assessed by methacholine challenge test, serum IgE levels, atopy, and atopic dermatitis. -
序列相似性
Belongs to the glycosyl hydrolase 18 family. -
翻译后修饰
Glycosylated. -
细胞定位
Secreted > extracellular space. - Information by UniProt
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数据库链接
- Entrez Gene: 1116 Human
- Omim: 601525 Human
- SwissProt: P36222 Human
- Unigene: 382202 Human
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别名
- 39 kDa synovial protein antibody
- ASRT7 antibody
- Cartilage glycoprotein 39 antibody
see all
图片
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All lanes : Anti-YKL-40/CHI3L1 antibody [EPR19078-157] (ab255297) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : CHI3L1 knockout THP-1 cell lysate
Lane 3 : U-87 MG cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 43 kDa
Observed band size: 37,40 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-YKL-40/CHI3L1 antibody [EPR19078-157] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab255297 was shown to bind specifically to YKL-40/CHI3L1. A band was observed at 37/40 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CHI3L1 knockout cell line ab280038 (knockout cell lysate ab280097). To generate this image, wild-type and CHI3L1 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling YKL-40/CHI3L1 with ab255297 at 1/250 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on neutrophils of human spleen is observed.
The section was incubated with ab255297 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution) for 20mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanolpermeabilized THP-1 (human monocytic leukemia cell line) cells labelling YKL-40/CHI3L1 with ab255297 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730, Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-YKL-40/CHI3L1 antibody [EPR19078-157] (ab255297) at 1/1000 dilution
Lane 1 : Human spleen tissue lysate
Lane 2 : THP-1 (Human monocytic leukemia monocyte) whole cell lysate
Lane 3 : U-87 MG ( Human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lane 4 : MG-63 (Human osteosarcoma fibroblast) whole cell lysate
Lane 5 : Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 6 : 293T (Human embryonic kidney epithelial cell) whole cell lysate
Lane 7 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 43 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?
Additional bands at: 27 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 60 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST.
The expression profile observed in 293T and Jurkat are consistent with the literature (PMID: 19767768,PMID: 16472595).
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling YKL-40/CHI3L1 with ab255297 at 1/250 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on macrophages in human tonsil germinal center is observed. The section was incubated with ab255297 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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All lanes : Anti-YKL-40/CHI3L1 antibody [EPR19078-157] (ab255297) at 1/1000 dilution
Lane 1 : Human spleen lysate
Lane 2 : THP-1 (human monocytic leukemia cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 43 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 minutes
The molecular weight observed is consistent with what has been described in the literature (PMID 19414556).
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (8)
ab255297 被引用在 8 文献中.
- Si Y et al. Inflammatory biomarkers, angiogenesis and lymphangiogenesis in epicardial adipose tissue correlate with coronary artery disease. Sci Rep 13:2831 (2023). PubMed: 36805000
- Zhao Y et al. Plasma proteome profiling using tandem mass tag labeling technology reveals potential biomarkers for Parkinson's disease: a preliminary study. Proteomics Clin Appl 16:e2100010 (2022). PubMed: 34791804
- Tu J et al. Single-Cell Transcriptome Profiling Reveals Multicellular Ecosystem of Nucleus Pulposus during Degeneration Progression. Adv Sci (Weinh) 9:e2103631 (2022). PubMed: 34825784
- Cai J et al. Increased Levels of CHI3L1 and HA Are Associated With Higher Occurrence of Liver Damage in Patients With Obstructive Sleep Apnea. Front Med (Lausanne) 9:854570 (2022). PubMed: 35280860
- Wang LJ et al. Tumor-associated macrophages related signature in glioma. Aging (Albany NY) 14:2720-2735 (2022). PubMed: 35332109
- Zhang W et al. SIRT1 inactivation switches reactive astrocytes to an antiinflammatory phenotype in CNS autoimmunity. J Clin Invest 132:N/A (2022). PubMed: 36136587
- Blanco-Palmero VA et al. Increased YKL-40 but Not C-Reactive Protein Levels in Patients with Alzheimer's Disease. Biomedicines 9:N/A (2021). PubMed: 34572280
- Hao H et al. LncRNA KCNQ1OT1 Promotes Proliferation, Invasion and Metastasis of Prostate Cancer by Regulating miR-211-5p/CHI3L1 Pathway. Onco Targets Ther 14:1659-1671 (2021). PubMed: 33688211