Anti-Smith Antigen抗体[Y12] (ab3138)

概述

  • 产品名称Anti-Smith Antigen抗体[Y12]
    参阅全部 Smith Antigen 一抗
  • 描述
    小鼠单克隆抗体[Y12] to Smith Antigen
  • 经测试应用适用于: Electron Microscopy, IHC-P, WB, ICC/IF, IPmore details
  • 种属反应性
    与反应: Rat, Human, Pig, Drosophila melanogaster
    预测可用于: Rabbit
  • 阳性对照
    • LS174T cells. Tonsil.

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • 存储溶液pH: 7.40
    Constituents: 0.0268% PBS, 0.2% BSA
  • Concentration information loading...
  • 纯度Protein G purified
  • 克隆单克隆
  • 克隆编号Y12
  • 同种型IgG3
  • 轻链类型kappa
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab3138 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Electron Microscopy Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Do not perform antigen retrieval.
WB Use at an assay dependent concentration. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).

This antibody precipitates the small nuclear RNAs U1, U2, U4, U5, and U6 providing direct evidence that the Sm antigen resides on all of these RNA-protein complexes.

ICC/IF 1/250. (See Abreview). Fix with 3% PFA.
IP Use at an assay dependent concentration.

靶标

  • 相关性Smith Antigen is an autoantigen on a series of particles composed of RNA and protein. Unlike RNP, the sm antigen is resistant to RNase, but partially sensitive to trypsin. It is involved in the pathogenesis of Systemic lupus erythematosus (SLE).
  • 细胞定位Nuclear
  • 别名
    • Sm antigen antibody
    • Smith Antigen antibody

Anti-Smith Antigen antibody [Y12] 图像

  • ab3138 at 1/100 dilution staining human HeLa cells (neuroblastoma) by ICC/IF. The cells were paraformaldehyde fixed, permeabilised with 1% triton in PBS and then blocked with 0.1% normal goat serum prior to incubation with the antibody for 35 minutes. A goat anti-mouse TRITC was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-Smith Antigen antibody [Y12] (ab3138) at 1 µg/ml

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 25 kDa
    Observed band size : 28 + 31 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 15 kDa,62 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute
  • Smith Antigen was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Mouse monoclonal to Smith Antigen and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab3138.

    Secondary: Protein G-HRP at 1/500 dilution.

    Band: 28 and 31kDa: Smith Antigen.

  • IHC image of ab3138 staining in human normal cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab3138, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Anti-Smith Antigen antibody [Y12] (ab3138)参考文献

This product has been referenced in:
  • Okeyo-Owuor T  et al. U2AF1 mutations alter sequence specificity of pre-mRNA binding and splicing. Leukemia 29:909-17 (2015). Read more (PubMed: 25311244) »
  • Prescott AR  et al. Time-resolved quantitative proteomics implicates the core snRNP protein SmB together with SMN in neural trafficking. J Cell Sci 127:812-27 (2014). Read more (PubMed: 24357717) »

See all 15 Publications for this product

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Molecular weight of IgG is ˜150-160 kDa. Heavy chain 51 kDa each and light chain 25 kDa each

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice ...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application CLIP
Sample Human Cell lysate - whole cell (HEK 293T)
Specification HEK 293T
Type iCLIP
Irradiation Energy Level (mJ/cm2): 150
Wavelength (nm): 254
Positive control Anti-hnRNP C antibody (sc-15386) bound protein G dynabeads, 0.1 mg/ml (antibody/beads)
Negative control Protein G dynabeads without antibody bound
Immuno-precipitation step Other - Protein G dynabeads
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Abcam user community

Verified customer

提交于 Mar 21 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (neuroblastoma, HeLa)
Specification neuroblastoma, HeLa
Fixative Paraformaldehyde
Blocking step Other as blocking agent for 30 minute(s) · Concentration: 0.1
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Abcam user community

Verified customer

提交于 Jul 24 2006

We don't have any information about this antibody being used in confocal microscopy; for Electron microscopy / Immunofluorescence it is recommended to use the antibody at an assay dependent concentration.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"