Anti-PMP70抗体(ab3421)

Anti-PMP70 antibody 图像

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  Western blot - Anti-PMP70 antibody (ab3421)
  All lanes : Anti-PMP70 antibody (ab3421) at 1/1000 dilution
  
  Lane 1 : Mouse lung cell lysate
  Lane 2 : Mouse liver cell lysate
  Lane 3 : HepG2 cell lysate
  
  Lysates/proteins at 25 µg per lane.
  
  
  Observed band size : 70 kDa (why is the actual band size different from the predicted?)
  
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PMP70 antibody - Peroxisomal Membrane Marker (ab3421)
  ab3421 (2µg/ml) staining PMP70 in human duodenum using an automated system (DAKO Autostainer Plus). Using this protocol there is apical cytoplasmic staining and staining of the endoplasmic reticulum in the epithelium.
  Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  
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  Immunocytochemistry/ Immunofluorescence - PMP70 antibody - Peroxisomal Membrane Marker (ab3421)

  ICC/IF image of ab3421 stained human HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3421, 1 µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor^® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in Hek293, HepG2 and MCF7 cells.

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  Flow Cytometry - Anti-PMP70 antibody (ab3421)

  Flow cytometry analysis of PMP70 in 293T cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10⁶ cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab3421 at a dilution of 0.5 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

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  Western blot - Anti-PMP70 antibody - Peroxisomal Membrane Marker (ab3421)
  All lanes : Anti-PMP70 antibody (ab3421) at 1/500 dilution
  
  Lane 1 : Rat kidney whole cell extracts at 20 µg
  Lane 2 : Rat liver whole cell extracts at 20 µg
  Lane 3 : Mouse lung whole cell extracts at 20 µg
  Lane 4 : A431 whole cell extracts at 10 µg
  Lane 5 : U2OS whole cell extracts at 10 µg
  
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  Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)

  Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in HMVEC Cells.

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  Flow Cytometry - Anti-PMP70 antibody (ab3421)

  Flow cytometry analysis of PMP70 in HepG2 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10⁶ cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab3421 at a dilution of 0.5 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

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  Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)

  Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in NS-1 Cells.

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  Flow Cytometry - Anti-PMP70 antibody (ab3421)

  Flow cytometry analysis of PMP70 in NIH-3T3 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10⁶ cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab3421 at a dilution of 0.5 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

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  Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)

  Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in p19 Cells.

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  Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)

  Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in A549 Cells.