Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790)

2 product images

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  Western blot - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175780)

  This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Milk before being incubated with Anti-alpha Tubulin antibody - Microtubule Marker ab18251 overnight at 4°C. Antibody binding was detected using ab175780 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

  All lanes: Western blot - Anti-alpha Tubulin antibody - Microtubule Marker (Anti-alpha Tubulin antibody - Microtubule Marker ab18251) at 1 µg/mL

  All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Western blot - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175780) at 1/10000 dilution

  Predicted band size: 36 kDa

  Observed band size: 50 kDa

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  ELISA - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175780)

  Cross-reactivity of the polyclonal secondary antibody Donkey Anti-Rabbit IgG H&L ab182020 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 μg/ml (50 μl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. Donkey Anti-Rabbit IgG H&L ab182020 was then added starting at 1 μg/ml and gradually diluted 1/4 (50 μl/well), followed by incubation for 2h. For the detection Goat anti-Donkey IgG H&L (HRP) (Goat Anti-Donkey IgG H&L (HRP) ab6988) was used at 1/20,000 dilution (50 μl/well), followed by incubation for 1h at RT.
  

  For the batch tested, Donkey Anti-Rabbit IgG H&L ab182020 showed a cross-reactivity below 2% towards human IgG, mouse IgG, rat IgG, goat IgG and chicken IgY.

  This data was developed using the unconjugated antibody (Donkey Anti-Rabbit IgG H&L ab182020).