小鼠单克隆抗体SB62a Anti-Rabbit IgG light chain (HRP) (ab99697)

概述

  • 产品名称小鼠单克隆抗体SB62a Anti-Rabbit IgG light chain (HRP)
    参阅全部 IgG Light chain 二抗
  • 靶标种属Rabbit
  • 特异性ab99697 reacts with the light chain of native Rabbit IgG primary antibodies and does not bind to the reduced and denatured Rabbit IgG heavy chain band (50kD).
  • 经测试应用适用于: ELISA, WBmore details
  • 免疫原

    Rabbit Light Chain

  • 偶联物HRP

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Store at +4°C.
  • 存储溶液Preservative: None
    Constituents: 50% PBS, 50% Glycerol, pH 7.4
  • Concentration information loading...
  • 纯度Protein A purified
  • 克隆单克隆
  • 克隆编号SB62a
  • 同种型IgG2b
  • 轻链类型lambda
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab99697 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
ELISA 1/4000 - 1/8000.
WB Use at an assay dependent concentration.

Note: This antibody is not recommended for sensitive or quantitative detection of the reduced and denatured Rabbit Light Chains on Western Blots.

Mouse monoclonal SB62a Anti-Rabbit IgG light chain (HRP) 图像

  • IHC image of beta actin staining in human colon formalin fixed paraffin embedded tissue section*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab8227, 3µg/ml overnight at +4°C. An HRP-conjugated secondary (ab99697, 1/8000 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.

    The inset negative control image is secondary-only at 1/4000 dilution.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Western blot showing that Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) specifically binds to light-chains


    Left blot: Coomassie stain showing the rabbit heavy (~50kDa) and light-chains (~25 kDa)

    Lane 1: 10 µg rabbit monoclonal was loaded
    Lane 2: 5 µg rabbit monoclonal was loaded
    Lane 3: 2 µg rabbit monoclonal was loaded

    Right blot: WB using Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) as secondary antibody at 1/10,000

    Lane 1: 1 µg rabbit monoclonal was loaded
    Lane 2: 0.5 µg rabbit monoclonal was loaded
    Lane 3: 0.25 µg rabbit monoclonal was loaded

  • All lanes : Anti-beta Actin antibody - Loading Control (ab8227) at 1 µg/ml

    Lanes 1-6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1 - 2 : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) at 1/2000 dilution
    Lanes 3 - 4 : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) at 1/10000 dilution
    Lanes 5 - 6 : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

  • LAP2 alpha was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to LAP2 alpha (ab5162)and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extract diluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab5162. Secodary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Bands: 83kDa: LAP2 alpha.
  • Catalase was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Catalase (ab1877) and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extract diluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab1877. Secodary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697). Bands: 60kDa: Catalase
  • Rad21 was immunoprecipitated using 0.5mg Hela whole cell extract, 5ug of Rabbit polyclonal to Rad21and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extractdiluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab42478. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) .. Band: 100kDa: Rad21; Non specific - 50kDa: We are unsure as to the identity of this extra band. Although the predicted band size is 75kDa based on Swiss-prot data, a band of 120kDa has been previously observed. Mol Cell Biol. 2002 Dec;22(23):8267-77 PMID: 12417729. Th
  • Lane 1: Rabbit IgG loaded at 1ug/well
    ab99697 diluted to 1:2000

Mouse monoclonal SB62a Anti-Rabbit IgG light chain (HRP) (ab99697)参考文献

This product has been referenced in:
  • Yang J  et al. PREX2 promotes the proliferation, invasion and migration of pancreatic cancer cells by modulating the PI3K signaling pathway. Oncol Lett 12:1139-1143 (2016). WB . Read more (PubMed: 27446408) »

See 1 Publication for this product

Product Wall

Application Immunoprecipitation
My protein of interest is slightly larger than heavy chain making the use of usual (H+L) secondaries difficult for WB following immunoprecipitation. The two images exemplify the problem. The first is using a secondary that detects both heavy and light chain. Heavy chain band covers my protein of interest. The second image was obtained after stripping the membrane and repeating the WB, this time using ab99697. Now the heavy chain band in the IP lane has disappeared and is faintly existent from the previous experiment in the IgG lane.

Good sensitivity as well. This example is 1:5000, but now I am using 1:10,000.

Excellent!
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Verified customer

提交于 Jul 17 2014

The has been tested to detect their specific proteins by Western blot under reducing and denatured conditions. Therefore they can recognise their targets in denatured conditions.

This ab appearsto be exactly what I assumed we did not have. The data does suggest that it will solve your problem, if your samples are reduced and denatured.

The rabbit light chain-specific antibody ab99697 should be suitable for your Western blot and I wouldn't expect it to cross-react with the reduced denatured mouse IgG heavy chain, however this antibody has not been tested for reactivity with mouse IgG ...

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