重组Anti-galectin 9/Gal-9抗体[EPR22214] (ab227046)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22214] to galectin 9/Gal-9
- Suitable for: ICC/IF, Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-galectin 9/Gal-9抗体[EPR22214]
参阅全部 galectin 9/Gal-9 一抗 -
描述
兔单克隆抗体[EPR22214] to galectin 9/Gal-9 -
宿主
Rabbit -
特异性
This antibody is not recommended for mouse and rat in IHC.
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经测试应用
适用于: ICC/IF, Flow Cyt (Intra), WB, IHC-Pmore details
不适用于: IP -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- IHC-P: Human liver and colon tissue. WB: U937, HL-60 and THP-1 whole cell lysates. ICC/IF: U937 cells. Flow Cyt (intra): U937 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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR22214 -
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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Immunohistochemistry reagents
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab227046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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ICC/IF |
1/250.
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Flow Cyt (Intra) |
1/500.
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WB |
1/1000. Predicted molecular weight: 40 kDa.
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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说明 |
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ICC/IF
1/250. |
Flow Cyt (Intra)
1/500. |
WB
1/1000. Predicted molecular weight: 40 kDa. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
靶标
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功能
Binds galactosides. Has high affinity for the Forssman pentasaccharide. May play a role in thymocyte-epithelial interactions relevant to the biology of the thymus. Inhibits cell proliferation. It is a ligand for HAVCR2/TIM3. Induces T-helper type 1 lymphocyte (Th1) death. Isoform Short acts as an eosinophil chemoattractant. -
组织特异性
Peripheral blood leukocytes and lymphatic tissues. Overexpressed in Hodgkin disease tissue. -
序列相似性
Contains 2 galectin domains. -
结构域
Contains two homologous but distinct carbohydrate-binding domains. -
细胞定位
Cytoplasm. Secreted. May also be secreted by a non-classical secretory pathway. - Information by UniProt
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数据库链接
- Entrez Gene: 3965 Human
- Omim: 601879 Human
- SwissProt: O00182 Human
- Unigene: 81337 Human
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别名
- 36 kDa beta-galactoside-binding lectin antibody
- Ecalectin antibody
- Gal-9 antibody
see all
图片
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All lanes : Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : LGALS9 knockout THP-1 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 40 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab227046 observed at 35 kDa. Red - loading control Mouse anti Histone H3 observed at 18 kDa.
ab227046 was shown to react with galectin 9/Gal-9 in wild-type THP-1 cells in Western blot with loss of signal observed in LGALS9 knockout cell line ab269505 (knockout cell lysate ab269667). Wild-type THP-1 and LGALS9 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab227046 and Mouse anti Histone H3 overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Immunocytochemistry analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Cell line U937 (human histiocytic lymphoma cell line) cells labeling galectin 9/Gal-9 with ab227046 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).Confocal image showing nuclear and cytoplasmic staining in U937 cell line.
Negative control: PANC-1.
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All lanes : Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046) at 1/500 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : Galectin 9 knockout A549 cell lysate
Lane 3 : U-937 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 40 kDa
Observed band size: 34-39 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab227046 observed at 34-39 kDa. Red - loading control ab8245 observed at 36 kDa.
ab227046 Anti-galectin 9/Gal-9 antibody [EPR22214] was shown to specifically react with galectin 9/Gal-9 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266923 (knockout cell lysate ab256976) was used. Wild-type and galectin 9/Gal-9 knockout samples were subjected to SDS-PAGE. ab227046 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemical analysis of paraffin-embedded human liver tissue labeling galectin 9/Gal-9 with ab227046 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear and cytoplasmic staining on Kupffer cells of human liver (PMID: 20209097, PMID: 18202194) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
All lanes : Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046) at 1/1000 dilution
Lane 1 : U937 (human histiocytic lymphoma monocyte), whole cell lysate
Lane 2 : HL-60 (human Acute Promyelocytic Leukemia promyeloblast), whole cell lysate
Lane 3 : MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate
Lane 4 : PANC-1 (human pancreatic epithelioid carcinoma epithelial cell), 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: 40 kDa
Observed band size: 34-39 kDa why is the actual band size different from the predicted?
Additional bands at: 13-15 kDa (possible degradation product)
Exposure time: 59 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression of the isoforms observed is consistent with what has been described in the literature (PMID 11886844, PMID: 22805533).
Degraded fragments (13-15 kDa) have also been descried (PMID: 15811318).
Negative control: PANC-1.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized PANC-1 (human pancreatic epithelial cancinoma cell line) (Left) / U937 (human histiocytic lymphoma cell line) (Right) cell line labeling galectin 9/Gal-9 with ab227046 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled 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.
Negative Control - PANC-1 cells.
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling galectin 9/Gal-9 with ab227046 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear and cytoplasmic staining on epithelial cells of human colon (PMID: 18202194) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney) cells labeling galectin 9/Gal-9 with ab227046 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing no staining in HEK-293T cells
Negative control: HEK-293T (PMID: 11698107). The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of the primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
All lanes : Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046) at 1/1000 dilution
Lane 1 : THP-1 (human monocytic leukemia monocyte), whole cell lysate
Lane 2 : HEK-293T (human embryonic kidney epithelial cell), 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: 40 kDa
Observed band size: 34-39 kDa why is the actual band size different from the predicted?
Additional bands at: 13-15 kDa (possible degradation product)
Exposure time: 59 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression of the isoforms observed is consistent with what has been described in the literature (PMID 11886844, PMID: 22805533).
Degraded fragments (13-15 kDa) has also been described (PMID: 15811318).
Negative control: HEK-293T (PMID: 24333696).
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 (4)
ab227046 被引用在 4 文献中.
- Liu X et al. Single-cell transcriptomics links malignant T cells to the tumor immune landscape in cutaneous T cell lymphoma. Nat Commun 13:1158 (2022). PubMed: 35241665
- Peng Q et al. Galectin-9/Tim-3 pathway mediates dopaminergic neurodegeneration in MPTP-induced mouse model of Parkinson's disease. Front Mol Neurosci 15:1046992 (2022). PubMed: 36479526
- Cao L et al. Prognostic Role of Immune Checkpoint Regulators in Cholangiocarcinoma: A Pilot Study. J Clin Med 10:N/A (2021). PubMed: 34069452
- Wu X et al. Sensing of mycobacterial arabinogalactan by galectin-9 exacerbates mycobacterial infection. EMBO Rep 22:e51678 (2021). PubMed: 33987949