IHC analysis for PCNA, pAKT, and pSTAT3 proteins was performed on formalin fixed, paraffin embedded orthotopic xenograft tumors (5 micron sections). Briefly, the tumor tissues were deparaffinized, rehydrated, treated with 0.3% hydrogen peroxide and processed for antigen retrieval using a heat-induced technique. Following blocking with background sniper (Biocare Medical, Concord, CA), the samples were processed for staining with PCNA, pAKT, and pSTAT3 antibodies (Cell Signaling Technologies, Danvers, MA). The expression of these proteins was detected using a MACH 4 Universal HRP Polymer detection kit (Biocare Medical) and 3, 9-diaminobenzidine (DAB substrate kit, Vector Laboratories, Burlingame, CA). The slides were counterstained with hematoxylin, dehydrated, mounted with VectaMount (Vector Laboratories) and visualized using an Olympus BX 41 Microscope (Olympus Corporation, Tokyo, Japan).
Mach 4 universal hrp polymer detection kit
Manufactured by Biocare Medical
Sourced in Canada
The MACH 4 Universal HRP Polymer detection kit is a reagent designed for the immunohistochemical detection of antigens in tissue sections. The kit utilizes a horseradish peroxidase (HRP) polymer detection system to amplify the signal, providing enhanced sensitivity for the visualization of target proteins.
Automatically generated - may contain errors
Lab products found in correlation
Background sniper, by Biocare Medical (4 mentions)
Vectamount, by Vector Laboratories (4 mentions)
Bx41 microscope, by Olympus (4 mentions)
3 39 diaminobenzidine dab substrate kit, by Vector Laboratories (2 mentions)
Pstat3 antibodies, by Cell Signaling Technology (1 mentions)
Proliferating cell nuclear antigen (pcna), by Cell Signaling Technology (1 mentions)
Biocare kit, by Biocare Medical (1 mentions)
Dab dab substrate kit, by Vector Laboratories (1 mentions)
Betazoid dab chromogen kit, by Biocare Medical (1 mentions)
Diva decloaker 10x, by Biocare Medical (1 mentions)
Ab32420, by Abcam (1 mentions)
Ab16667, by Abcam (1 mentions)
7 protocols using mach 4 universal hrp polymer detection kit
1
Immunohistochemical Analysis of Tumor Proteins
2
Immunohistochemistry Characterization of Xenograft Tumors
3
Immunohistochemical Analysis of Cell Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A heat-induced technique was practiced to deparaffinize, rehydrate, and treat the paraffin-embedded tissues by 0.3% hydrogen peroxide for antigen retrieval. Then for staining, the samples were treated with Ki-67, E-cadherin and vimentin antibodies (Abcam, Cambridge MA) and background sniper (Biocare Medical, Concord, CA). MACH 4 Universal HRP Polymer detection kit (Biocare Medical) and 3,39-diaminobenzidine (DAB substrate kit, Vector Laboratories, Burlingame, CA) were used for measuring the proteins expression. The hematoxylin, dehydrated, mounted with VectaMount (Vector Laboratories) were employed to counter stain the slides, and Olympus BX 41 Microscope (Olympus Corporation, Japan) for visualization.
4
Immunohistochemical Analysis of Oncogenic HPV Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IHC analysis for HPV E6/E7 proteins, Ki67 and PTEN on formalin fixed, paraffin embedded orthotopic mouse tumors (5 micron sections) was performed. Briefly, the tumor tissues were deparaffinized, rehydrated, treated with 0.3% hydrogen peroxide and processed for antigen retrieval using a heat-induced technique. Following blocking with background sniper (Biocare Medical, Concord, CA), the samples were processed for staining with E6 (Abcam, Cambridge MA), E7 (Invitrogen), Ki67 and PTEN antibodies (Cell Signaling Technologies). The expression of these proteins was detected using a MACH 4 Universal HRP Polymer detection kit (Biocare Medical) and 3,39-diaminobenzidine (DAB substrate kit, Vector Laboratories, Burlingame, CA). The slides were counterstained with hematoxylin, dehydrated, mounted with VectaMount (Vector Laboratories) and visualized using an Olympus BX 41 Microscope (Olympus Corporation, Japan).
5
Immunohistochemical Analysis of PCNA and MUC13
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IHC analysis for PCNA and MUC13 proteins was performed on formalin-fixed, paraffin-embedded xenograft tumors (5-micron sections) as described previously [20 (link)]. Briefly, the tumor tissues were deparaffinized, rehydrated, treated with 0.3 percent hydrogen peroxide, and then processed for antigen retrieval while using a heat-induced technique. The samples were processed for staining with PCNA and MUC13 antibodies after blocking with background sniper (BioCare Medical, Concord, CA, USA). The expression of these proteins was detected while using a MACH 4 Universal HRP Polymer Detection Kit (BioCare Medical) and 3,9-diaminobenzidine (DAB Substrate Kit, Vector Laboratories, Burlingame, CA, USA). The slides were counter-stained with hematoxylin, dehydrated, mounted with VectaMount (Vector Laboratories), and visualized using an Olympus BX 41 microscope (Olympus Corporation, Tokyo, Japan).
6
Immunohistochemical Analysis of CCL4 in CRC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin-fixed and paraffin-embedded CRC tissues from patients with CRC were obtained from the Department of Pathology of the County Hospital Ryhov, Jönköping, Sweden. Among the eight patients, six had tumors localized in the colon and two in the rectum; these were classified as stage I in two cases, stage II in two cases, stage III in two cases, and stage IV in two cases. The staining was performed on 3.5 μm sections of paraffin-embedded tissue samples to detect the CCL4 protein expression. In brief, antigen retrieval was finished by cooking at 110°C for 52 min in Diva Decloaker, 10X (Biocare Medical, Concord, CA, United States). The slides were treated with hydrogen peroxide for 5 min to prevent the occurrence of endogenous peroxidase, which might alter the interpretation of the color. Primary rabbit polyclonal CCL4 antibody against human CCL4 (Abcam, Tokyo, Japan) was used at a dilution of 1:300. The antibody was applied to the tissue sections, which were then incubated for 30 min at room temperature. The MACH 4 Universal HRP-Polymer Detection kit (Biocare Medical) was used, and the reaction was visualized using Betazoid DAB Chromogen Kit (Biocare Medical). Human kidney tissue was used as a positive control for CCL4 expression and rabbit IgG, polyclonal isotype control (Abcam) was used as a negative control and included along with each patient tissue section.
7
Immunohistochemical Analysis of Ki67 and ATM
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FFPE Tissue sections were deparaffinized and rehydrated using standard procedures. Heat-induced epitope retrieval was carried out in a BioCare Medical (Concord, CA, USA) pressure cooker (Decloaking ChamberTM NxGen) with 10mM citrate buffer (pH 6.0) at 110°C for 12 min. Samples were incubated with a rabbit anti-Ki67 antibody (ab16667, Abcam, UK, diluted 1:100), or a rabbit anti-ATM antibody (ab32420, Abcam, UK, diluted 1:200) respectively 1 hour at RT or overnight at 4°C in a moist chamber. The signal was revealed with MACH 4 Universal HRP-Polymer Detection Kit (BioCare) according to the manufacturer's instructions. Purified rabbit IgG (02–6102, Life Technologies, Switzerland) was used as a negative control. Stained sections were photographed with a Zeiss Axio Vert.A1 microscope. Images were imported with AxioVision v. 4.8.2.0.
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!