Anti-PMP70 antibody [EPR5614]

6 product images

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  Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)

  Lanes 1- 2: Merged signal (red and green). Green - ab109448 observed at 75 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
  

  ab109448 was shown to react with PMP70 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265294 (knockout cell lysate ab257134) was used. Wild-type HeLa and ABCD3 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109448 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (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.

  All lanes: Western blot - Anti-PMP70 antibody [EPR5614] (ab109448) at 1/1000 dilution

  Lane 1: Wild-type HeLa cell lysate at 40 µg

  Lane 2: ABCD3 knockout HeLa cell lysate at 40 µg

  Performed under reducing conditions.

  Predicted band size: 75 kDa

  Observed band size: 75 kDa

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  Flow Cytometry (Intracellular) - Anti-PMP70 antibody [EPR5614] (ab109448)

  ab109448 at 1/100 dilution staining PMP70 in permeabilized HepG2 cells by intracellular flow cytometry (shown in red). Rabbit IgG negative control (shown in green).

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  Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)

  Lanes 1 - 4: Merged signal (red and green). Green - ab109448 observed at 75 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.
  

  ab109448 was shown to specifically react with PMP70 in wild-type HAP1 cells as signal was lost in ABCD3 (PMP70) knockout cells. Wild-type and ABCD3 (PMP70) knockout samples were subjected to SDS-PAGE. ab109448 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-PMP70 antibody [EPR5614] (ab109448) at 1/1000 dilution

  Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

  Lane 2: ABCD3 (PMP70) knockout HAP1 whole cell lysate at 20 µg

  Lane 3: A431 whole cell lysate at 20 µg

  Lane 4: HEPG2 whole cell lysate at 20 µg

  Predicted band size: 75 kDa

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  Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)

  All lanes: Western blot - Anti-PMP70 antibody [EPR5614] (ab109448) at 1/1000 dilution

  Lane 1: 293T cell lysate at 10 µg

  Lane 2: SH SY5Y cell lysate at 10 µg

  Lane 3: Caco2 cell lysate at 10 µg

  Lane 4: HepG2 cell lysate at 10 µg

  Predicted band size: 75 kDa

  Observed band size: 70 kDa

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  OI-RD Scanning - Anti-PMP70 antibody [EPR5614] (ab109448)

  We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
  Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

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  Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody [EPR5614] (ab109448)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling PMP70 with primary antibody anti-PMP70 (ab109448) at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody at 1/1000 dilution (2.0 μg/mL). Confocal image showing cytoplasmic staining on HeLa cells. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) was used to counterstain tubulin at 1/200 dilution (2.5 μg/mL). The nuclear counter stain is DAPI (blue).
  The secondary antibody only control is : Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (2.0 μg/mL).