Formalin-fixed paraffin-embedded tissues (FFPE) and tumor microarray (TMA) sections (5 µm thickness) were deparaffinized in xylene and rehydrated in descending alcohols, followed by blocking of endogenous peroxidase in 3% hydrogen peroxide solution (30 min) and heat-induced epitope retrieval in citrate buffer (95 °C, 5 min). Sections were incubated with CD206 (1:100, Clone: C-10, Santa Cruz), iNOS (1:100, Clone: C-11, Santa Cruz), CD16b (1:100, Clone: CLB-gran11.5, BD Biosciences), Ly6G (1:100, Clone: 1A8, BioLegend), p-Smad3 (1:100, Clone: 1D9, Santa Cruz), p-Smad3 (1:200, 600-401-919, Rockland) antibodies overnight (4 °C), sections were incubated in polymer-HRP conjugated secondary antibody (Dako) for 2 h at room temperature, followed by DAB (Thermo-fisher) or Opal-520, 570, 650, 690 TSA dye (Akoya biosciences) development. DAB-stained section images were captured by the Nikon Ni-U Light Microscope and analyzed using Image J analysis software; Opal TSA-stained sections were captured on the Mantra quantitative pathology workstation (Akoya Biosciences) and analyzed by inForm image analysis software 2.6 (Akoya Biosciences) as per our previous studies18 (link),51 (link).
Polymer hrp conjugated secondary antibody
Manufactured by Agilent Technologies
The Polymer-HRP conjugated secondary antibody is a laboratory reagent designed to aid in the detection and visualization of target proteins in various immunoassay applications. It consists of a polymer backbone conjugated with multiple horseradish peroxidase (HRP) enzymes. The HRP enzymes can catalyze a colorimetric reaction, allowing for the amplification and detection of the target protein signal.
Automatically generated - may contain errors
Lab products found in correlation
P smad3, by Santa Cruz Biotechnology (1 mentions)
Cd206, by Santa Cruz Biotechnology (1 mentions)
3 3 diaminobenzidine (dab), by Thermo Fisher Scientific (1 mentions)
3 3 diaminobenzidine (dab), by Nikon (1 mentions)
Ni u light microscope, by Nikon (1 mentions)
Mantra quantitative pathology workstation, by Akoya Biosciences (1 mentions)
Psmad3, by Rockland Immunochemicals (1 mentions)
Cd16b, by BD (1 mentions)
Pepsin, by Agilent Technologies (1 mentions)
Blocking solution, by Agilent Technologies (1 mentions)
3 3 diaminobenzidine substrate chromogen system, by Agilent Technologies (1 mentions)
Ix71 inverted microscope, by Olympus (1 mentions)
2 protocols using polymer hrp conjugated secondary antibody
1
Immunohistochemical Analysis of FFPE Tissues
2
Histological Analysis of Tissue Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mice were killed after the experimental treatments. Soon thereafter, 4-mm-thick sections were cut, fixed in formalin and embedded in paraffin before being dewaxed and rehydrated as previously described.27 (link) For hematoxylin and eosin (H&E) staining, the sections were incubated with H&E. For IHC, the sections were incubated in a 3% hydrogen peroxide/methanol solution. Antigen retrieval was performed by incubation with 0.25% pepsin (Dako, Carpinteria, CA, USA). The sections were then blocked in a blocking solution (Dako) and incubated overnight with the indicated primary antibody (ABcam, Cambridge, UK) at 4 °C. After washing with TBST, the sections were incubated with a polymer-HRP-conjugated secondary antibody (Dako). Following hematoxylin staining and bluing (1% HCl in 70% ethanol), a 3,3′-diaminobenzidine Substrate Chromogen System (Dako) was used to detect antibody binding, and the stained sections were examined with an Olympus IX71 inverted microscope (Olympus Optical Co. Ltd.).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!