Donkey Anti-Mouse IgG H&L (HRP)

4 product images

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)

  IHC image of Histone H4 staining in a section of formalin-fixed paraffin-embedded normal human colon*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade ab31830 at 0.5ug/ml. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature.

  An HRP-conjugated secondary (ab205724, 1/2000 dilution) was used for 1hr at room temperature.

  The section was counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)

  IHC image of alpha tubulin staining in a section of formalin-fixed paraffin-embedded normal human colon*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 0.5ug/ml.

  An HRP-conjugated secondary (ab205725, 1/2000 dilution) was used for 1hr at room temperature. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

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

• 

  Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)

  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 2% Bovine Serum Albumin before being incubated with Anti-beta Actin antibody [mAbcam 8226] - Loading Control ab8226 overnight at 4°C. Antibody binding was detected using ab205724, and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.

  All lanes: Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (Anti-beta Actin antibody [mAbcam 8226] - Loading Control ab8226) at 1 µg/mL

  Lane 1: Liver (Human) Tissue Lysate at 10 µg

  Lane 2: Liver (Mouse) Tissue Lysate at 10 µg

  Lane 3: Liver (Rat) Tissue Lysate at 10 µg

  Lane 4: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Lane 5: NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

  Lane 6: PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724) at 1/2000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 36 kDa

  Observed band size: 42 kDa

  Exposure time: 15s

• 

  Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)

  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 incubated overnight with 2% Bovine Serum Albumin at 4°C. Any non-specific background binding was assessed by incubating the membrane with only the secondary antibody (ab205724), and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.

  All lanes: No Primary Antibody

  Lane 1: Liver (Human) Tissue Lysate at 10 µg

  Lane 2: Liver (Mouse) Tissue Lysate at 10 µg

  Lane 3: Liver (Rat) Tissue Lysate at 10 µg

  Lane 4: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Lane 5: NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

  Lane 6: PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724) at 1/2000 dilution

  Performed under reducing conditions.

  Exposure time: 15s