Donkey Anti-Goat IgG H&L (HRP)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling GFAP with ab302644 at 1/1000 (0.998 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on human cerebrum. The section was incubated with ab302644 for 30 mins at room temperature, the 2nd antibody Donkey Anti-Goat IgG (HRP) ab205723 was used.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling MAP2 with ab302487 at 1/1000 (1.004 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on ganglion in the human colon. The section was incubated with ab302487 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling MAP2 with ab302487 at 1/1000 (1.004 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on human cerebrum. The section was incubated with ab302487 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labeling non-muscle Myosin IIB/MYH10 with ab300649 at 1/500 (1.96 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on human cerebellum. The section was incubated with ab300649 for 30 mins at room temperature, the 2nd antibody Donkey Anti-Goat IgG (HRP) ab205723 was used.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling MAP2 with ab302487 at 1/1000 (1.004 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on ganglion in the mouse colon. The section was incubated with ab302487 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling MAP2 with ab302487 at 1/1000 (1.004 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on rat cerebrum. The section was incubated with ab302487 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling GFAP with ab302644 at 1/1000 (0.998 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on mouse cerebrum. The section was incubated with ab302644 for 30 mins at room temperature, the 2nd antibody Donkey Anti-Goat IgG (HRP) ab205723 was used.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling non-muscle Myosin IIB/MYH10 with ab300649 at 1/500 (1.96 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on rat cerebellum. The section was incubated with ab300649 for 30 mins at room temperature, the 2nd antibody Donkey Anti-Goat IgG (HRP) ab205723 was used.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling non-muscle Myosin IIB/MYH10 with ab300649 at 1/500 (1.96 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on mouse cerebellum. The section was incubated with ab300649 for 30 mins at room temperature, the 2nd antibody Donkey Anti-Goat IgG (HRP) ab205723 was used.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling MAP2 with ab302487 at 1/1000 (1.004 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on ganglion in the rat colon. The section was incubated with ab302487 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling GFAP with ab302644 at 1/1000 (0.998 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on rat cerebrum. The section was incubated with ab302644 for 30 mins at room temperature, the 2nd antibody Donkey Anti-Goat IgG (HRP) ab205723 was used.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

Lab

Western blot - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

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 (ab205723), 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-Goat IgG H&L (HRP) (ab205723) at 1/2000 dilution

false

Exposure time: 5s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

IHC image of alpha tubulin staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*. 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 1ug/ml. ab98800, goat anti-mouse IgG, was then added as a secondary bridging antibody, at 1/250 dilution for 1h.

An HRP-conjugated secondary (ab205723, 1/2000 dilution) was used to detect the primary 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

• WB

Lab

Western blot - Donkey Anti-Goat IgG H&L (HRP) (AB205723)

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 ab8229 overnight at 4°C. Antibody binding was detected using ab205723, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-beta Actin antibody - Loading Control (<a href='/en-us/products/primary-antibodies/beta-actin-antibody-loading-control-ab8229'>ab8229</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-Goat IgG H&L (HRP) (ab205723) at 1/10000 dilution

Predicted band size: 36 kDa

Observed band size: 42 kDa

true

Exposure time: 30s