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

7 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

  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 Anti-beta Actin antibody 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

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  Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

  LAP2 alpha was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to LAP2 alpha (Anti-LAP2 alpha antibody 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 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with Anti-LAP2 alpha antibody ab5162. Secodary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
  Bands: 83kDa: LAP2 alpha.
  

  Secondary

  All lanes: Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

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  Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

  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

  Secondary

  All lanes: Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

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  Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

  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

  Secondary

  All lanes: Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

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  Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

  All lanes : Anti-beta Actin antibody - Loading Control (Anti-beta Actin antibody 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.

  Secondary

  All lanes: Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

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  Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

  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 70^oC; 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

  Secondary

  All lanes: Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

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  Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

  Lane 1: Rabbit IgG loaded at 1ug/well
  
  ab99697 diluted to 1:2000

  Secondary

  All lanes: Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)