Donkey Anti-Mouse IgG H&L (HRP)

• WB

Lab

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 ab8226 overnight at 4°C. Antibody binding was detected using ab205724, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (<a href='/ko/products/primary-antibodies/beta-actin-antibody-mabcam-8226-loading-control-ab8226'>ab8226</a>) 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

Predicted band size: 36 kDa

Observed band size: 42 kDa

true

Exposure time: 15s

• IHC-P

Lab

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 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

• IHC-P

Lab

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 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

• WB

Lab

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 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

false

Exposure time: 15s