IHC-P of ab88147 on human pancreas. antibody concentration 3 ug/ml.

This image was generated using the ascites version of the product.

All lanes : Anti-Collagen I antibody [3G3] (ab88147) at 1/1000 dilution

Lane 1 : Mouse skin
Lane 2 : Mouse placenta
Lane 3 : Mouse kidney
Lane 4 : Mouse liver

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : IR- goat anti-mouse (green) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 139 kDa

This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system under denaturing, reducing conditions. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with mouse anti-Collagen I antibody (88147; 1:1000) and the loading control rabbit anti-GAPDH antibody (ab9485; 1:10000) overnight at 4°C. Antibody binding was detected using infrared (IR) labelled goat anti-mouse (green; 1:10000) and IR-goat anti-rabbit (red; 1:10000) for 1 hour at room temperature before imaging.

This image was generated using the ascites version of the product.

IHC image of collagen I staining in a section of normal mouse placenta formalin-fixed paraffin-embedded. 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 ab88147 at 1/200 dilution. An HRP-conjugated secondary (Ab97040, 1/500 dilution) was used for 1hr at room temperature. DAB was used as the chromogen (10min at room temperature). The section was counterstained with haematoxylin and mounted with DPX. 

 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.

This image was generated using the ascites version of the product.

Anti-Collagen I antibody [3G3] (ab88147) at 1 µg/ml + immunogen (88 aa recombinant fragment with a proprietary tag of 26kDa) at 0.2 µg

Secondary
Goat anti-Mouse IgG at 1/5000 dilution

Predicted band size: 139 kDa

This image was generated using the ascites version of the product.

IHC image of Collagen I staining in normal human placenta formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab88147, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

 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

This image was generated using the ascites version of the product.

Anti-Collagen I antibody [3G3] (ab88147) at 1 µg/ml + NIH 3T3 cell lysate at 25 µg

Secondary
Goat anti-Mouse IgG at 1/5000 dilution

Predicted band size: 139 kDa

This image was generated using the ascites version of the product.

IHC image of Collagen I staining in human skin formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab88147, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

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.

This image was generated using the ascites version of the product.

Detection limit for ab88147 is 0.03 ng/ml as a capture antibody.

This image was generated using the ascites version of the product.