Kidney sections cut at 4 μm were deparaffinized to PBS. Antigen retrieval was performed by in citrate buffer (pH 6.0) for 15 min. Once cooled, sections were incubated with 3% H2O2 for 20 min to inactivate endogenous peroxidases (Dako EnVision+ System‐HRP‐DAB, K4010) and subsequently treated with 10% normal goat serum (Dako, UK) to reduce non‐specific absorption. Sections were subsequently incubated at 37°C for 1 h with the following primary antibodies: anti‐F4/80 (Abcam Cat# ab111101, RRID:AB_10859466) washed with PBS and then incubated at room temperature for 30 min with labelled polymer‐HRP antibody (Dako EnVision+ System, HRP‐DAB). Sections were developed in DAB chromogen solution, and the reaction stopped by immersion of sections in water. Counterstaining was performed with Harris haematoxylin before sections were dehydrated and mounted in DPX mounting medium.
Dako envision system hrp dab
Manufactured by Agilent Technologies
Sourced in United Kingdom, United States
The Dako Envision+ System-HRP DAB is a laboratory equipment product used for immunohistochemistry (IHC) applications. It is a polymer-based detection system that utilizes horseradish peroxidase (HRP) enzyme and 3,3'-diaminobenzidine (DAB) chromogen to visualize target antigens in tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Normal goat serum (ngs), by Agilent Technologies (2 mentions)
Target retrieval solution, by Agilent Technologies (2 mentions)
Mounting solution, by Electron Microscopy Sciences (2 mentions)
Meyer s hematoxylin, by Merck Group (2 mentions)
Protein blocking solution, by Agilent Technologies (2 mentions)
Protein block solution, by Agilent Technologies (1 mentions)
Histoclear solution, by National Diagnostics (1 mentions)
Paraformaldehyde solution, by Thermo Fisher Scientific (1 mentions)
Antibody diluent with background reducing components, by Agilent Technologies (1 mentions)
Protein block serum free solution, by Agilent Technologies (1 mentions)
Imager m2, by Zeiss (1 mentions)
Target retrieval solution ph 6, by Agilent Technologies (1 mentions)
Histoclear, by Electron Microscopy Sciences (1 mentions)
Anti α smooth muscle actin antibody, by Abcam (1 mentions)
Anti zeb1, by Abcam (1 mentions)
Ab15580, by Abcam (1 mentions)
Blocking solution, by PerkinElmer (1 mentions)
Peroxidase blocking solution, by Agilent Technologies (1 mentions)
Bx51 upright microscope, by Olympus (1 mentions)
Rabbit anti myeloperoxidase antibody, by Abcam (1 mentions)
11 protocols using dako envision system hrp dab
1
Macrophage Immunohistochemistry in Kidney Sections
2
Histological Analysis of Mouse Stomach
3
Immunohistochemical Staining of FFPE Tissues
4
Immunohistochemical Detection of Mutant p53
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IHC was performed as described previously29 (link) with minor modification. Briefly, slides were deparaffinised and rehydrated through series of graded ethanol solutions. Antigen retrieval was performed using a pressure cooker. The endogenous peroxidase activity was blocked by incubating the slides with 3% H2O2 for 30 min followed by incubation with protein blocking solution (Dako, Glostrup, Denmark) for 1 hr at room temperature. Next, the slides were at first incubated with the mutant p53 antibody in a humid chamber at 4 °C overnight, and were then incubated with the secondary rabbit IgG (Dako) antibody for 15 minutes at room temperature, and developed with Dako Envision+ System-HRP DAB (Dako). After counterstaining with Meyer’s Hematoxylin (Sigma-Aldrich), the slides were mounted with the mounting solution (Electron Microscopy Sciences, Hatfield, PA, USA).
5
Immunohistochemical Analysis of Tissue Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sections cut at 4 μm were dewaxed and deparaffinized to PBS. Antigen retrieval was performed by microwaving (700 W) sections in citrate buffer (pH 6.0) for 15 minutes. Once cooled, sections were incubated with 3% H2O2 for 20 minutes to inactivate endogenous peroxidises (Dako EnVision+ System‐HRP‐DAB, K4010) and subsequently treated with 10% normal goat serum (Dako, UK) to reduce nonspecific absorption. Sections were subsequently incubated at 37°C for 1 hour with the following primary antibodies: anti‐α‐smooth muscle actin antibody (1:400, cat#ab5694, Abcam, UK) or anti‐CD68 antibody ED1 (1:400; cat#MCA341R, AbD Serotec, UK), washed with PBS, and then incubated at room temperature for 30 minutes with labelled polymer‐HRP antibody (Dako EnVision+ System, HRP‐DAB). Sections were developed in DAB chromogen solution, and the reaction stopped by immersion of sections in water. Counterstaining was performed with Harris hematoxylin before sections were dehydrated and mounted in DPX mounting medium. Ten images per section were acquired with the NanoZoomer Digital Pathology Scanner in a double‐blinded manner. Quantification of staining was then performed using ImageJ analysis software.
6
Immunohistochemical Analysis of ZEB1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IHC analyses were performed as previously described with minor modification41 (link). Briefly, slides were deparaffinized and rehydrated through series of graded ethanol. Antigen retrieval was performed using a pressure cooker. Endogenous peroxidase activity was blocked by incubation with 3% H2O2 for 30 min then incubated with protein blocking solution (Dako, Glostrup, Denmark) for 1 hour at room temperature. Primary antibody was incubated in a humid chamber at 4 °C overnight, and then slides were incubated with secondary rabbit IgG (Dako) for 15 min at room temperature, and developed with Dako Envision + System-HRP DAB (Dako). Anti-ZEB1 (Abcam, Cambridge, UK; dilution ratio 1:2000) was purchased. After counterstaining with Meyer’s Hematoxylin (Sigma), slides were mounted with mounting solution (Electron Microscopy Sciences, Hatfield, PA, USA). Quantitation of ZEB1-positive cells was done and then statistical analyses were performed with JMP software. Student’s t-tests were used to analyze differences between means. Data are represented as mean ± SEM.
7
Immunohistochemical Staining of Paraffin-Embedded Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The paraffin‐embedded tissue sections were deparaffinized in xylene, rehydrated in ethanol, and immersed in citrate‐NaOH buffer (10 mmol L−1 sodium citrate, pH 6.0) at 121°C for 20 minutes. After retrieval of antigenicity, the nonspecific antibody reaction was blocked in blocking solution (Perkin Elmer Life Science), and the samples were incubated with antibodies against E‐cadherin (610181, BD Biosciences, 1:100) or Ki‐67 (ab15580, Abcam, 1:500). After the sections had been washed, the reacted antibodies were detected using the Dako EnVision + System/HRP (DAB) (DakoCytomation).
8
Paraffin Embedding and Immunohistochemistry of P2 Cells
9
Immunohistochemical Analysis of CCDC110 in Human Testis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The IHC experiment using human tissue samples was approved from Dankook
University Hospital IRB in 2006. The paraffin embedded tissue blocks of human
testis previously obtained from a patient in his 50s who was hospitalized after
a car accident were cut into 10-μm sections and placed on frosted glass
microscope slides. After removal of paraffin with xylene, the tissue sections
were dehydrated in a graded alcohol series. For the procedure of antigen
retrieval, the tissue sections were heated in a pressured chamber containing 10
mM sodium citrate buffer (pH 6.1) for 3 min. After blocking of endogenous
peroxide activity using 0.03% hydrogen peroxide, the sections were
incubated for 2 h with a primary antibody (1:1 to 1:2 diluted culture soup for
mAbs, 1:1,000 for polyclonal antibody) against CCDC110 at room temperature. The
samples were washed and then incubated with HRP-conjugated anti-mouse IgG (Dako
EnVision+system-HRP [DAB], Dako, Carpinteria, CA, USA) for 20 min at room
temperature. After washing, the chromogen was developed for 2 min. The tissue
sections were then counterstained with weak hematoxylin. The images of IHC was
obtained using Olympus BX51 upright microscope (Olympus, Tokyo, Japan) equipped
with digital camera.
University Hospital IRB in 2006. The paraffin embedded tissue blocks of human
testis previously obtained from a patient in his 50s who was hospitalized after
a car accident were cut into 10-μm sections and placed on frosted glass
microscope slides. After removal of paraffin with xylene, the tissue sections
were dehydrated in a graded alcohol series. For the procedure of antigen
retrieval, the tissue sections were heated in a pressured chamber containing 10
mM sodium citrate buffer (pH 6.1) for 3 min. After blocking of endogenous
peroxide activity using 0.03% hydrogen peroxide, the sections were
incubated for 2 h with a primary antibody (1:1 to 1:2 diluted culture soup for
mAbs, 1:1,000 for polyclonal antibody) against CCDC110 at room temperature. The
samples were washed and then incubated with HRP-conjugated anti-mouse IgG (Dako
EnVision+system-HRP [DAB], Dako, Carpinteria, CA, USA) for 20 min at room
temperature. After washing, the chromogen was developed for 2 min. The tissue
sections were then counterstained with weak hematoxylin. The images of IHC was
obtained using Olympus BX51 upright microscope (Olympus, Tokyo, Japan) equipped
with digital camera.
10
Immunohistochemical Staining of ERRFI1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue sections were mounted on the coated slides, deparaffinized with xylene, hydrated in serial solutions of alcohol, and heated in a pressure cooker containing 10 mmol/L sodium citrate (pH 6.0) for 3 min at full power for antigen retrieval. Endogenous peroxidase activity blocking was performed using 0.03% hydrogen peroxide containing sodium azide for 5 min. The sections were incubated at room temperature for 4 h with the rabbit polycloncal anti-ERRFI1 (Mig-6) antibody (1,50, ab198834, Abcam, Cambridge, United Kingdom). After washing, the samples were incubated in labelled polymer-HRP anti-mouse (Dako EnVision+system-HRP (DAB), Dako, Carpinteria, California, USA) for an additional 20 min at room temperature followed by additional washing. After rinsing, chromogen was developed for 2 min. The slides were then counterstained with Meyer’s hematoxylin, dehydrated, and coverslipped. Immunohistochemistry staining was scored to evaluate both intensity of the staining and proportion of stained tumor cells in each stained slide. The intensity was scored as 0 (negative), + 1 (mild), + 2 (moderate), + 3 (marked) and proportions were scored ranged from 0 to 100%.
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!