Immpact dab peroxidase hrp substrate

Manufactured by Vector Laboratories

Sourced in United States

ImmPACT DAB Peroxidase (HRP) Substrate is a chromogenic substrate used for the detection of horseradish peroxidase (HRP) in immunohistochemistry and other HRP-based applications. It produces a brown precipitate at the site of the HRP enzyme, allowing for visual identification of the target antigen.

Automatically generated - may contain errors

Lab products found in correlation

Vectastain elite abc hrp kit, by Vector Laboratories (5 mentions) Immpress hrp anti rabbit igg peroxidase polymer detection kit, by Vector Laboratories (5 mentions) Axioscan, by Zeiss (3 mentions) Vectastain abc hrp kit, by Vector Laboratories (3 mentions) Hematoxylin, by Vector Laboratories (3 mentions) Bloxall, by Vector Laboratories (2 mentions) Zen lite 2012, by Zeiss (2 mentions) Anti fapα, by R&D Systems (2 mentions) Anti goat horseradish peroxidase hrp, by Agilent Technologies (2 mentions) Anti fapα f11 24, by Thermo Fisher Scientific (2 mentions) Anti mouse igg2a tritc, by Southern Biotech (2 mentions) Anti mouse igg2b cy5, by Southern Biotech (2 mentions) Goat anti fitc alexa 488 antibody, by Thermo Fisher Scientific (2 mentions) Goat anti mouse igg1 fitc, by Southern Biotech (2 mentions) Anti pdpn nz 1, by Thermo Fisher Scientific (2 mentions) Thy 1a1, by R&D Systems (2 mentions) Lsm 510 confocal microscope, by Zeiss (2 mentions) Avidin biotin blocking kit, by Vector Laboratories (2 mentions) Rabbit anti cleaved caspase 3 antibody, by Cell Signaling Technology (2 mentions) Nanozoomer digital slide scanner, by Hamamatsu Photonics (2 mentions)

43 protocols using immpact dab peroxidase hrp substrate

Quantifying Angiogenesis in Fracture Healing

Check if the same lab product or an alternative is used in the 5 most similar protocols

Vascularity was analyzed at Day 5 and Day 14 for full fracture by quantifying blood vessels in high powered fields (20x) of the expanded periosteum that were 1 mm and 2 mm from the fracture site. For the stress fracture model, blood vessels in the entire expanded periosteal callus (area between cortical bone and skeletal muscle) was quantified at Day 3, Day 5, and Day 7. Vessels were visualized using immunohistochemistry with a rat monoclonal endomucin (clone eBioV.7C7, ebisocience; 1:400 dilution) antibody on paraffin sections. Continuous structures that were endomucin⁺ were counted as blood vessels. Sections underwent standard IHC paraffin protocols including deparaffinization in xylene and rehydration in graded ethanols. Antigen retrieval was done by proteinase K (5 min, room temp) with subsequent processing done according to the Vectastain Elite ABC HRP kit (Vector Labs; PK-6104). At least one negative control section was run per batch using the same procedures but with an isotype control antibody (Clone eBR2a, ebioscience; 1:400 dilution). Detection was done with ImmPact DAB Peroxidase (HRP) Substrate (Vector Labs; SK-4105). Sections were imaged at 20x on a Nanozoomer Slide Scanner (Hamamatsu) and analyzed by a blinded user to mouse ID and experimental group.

Buettmann E.G., McKenzie J.A., Migotsky N., Sykes D.A., Hu P., Yoneda S, & Silva M.J. (2019). VEGFA from Early Osteoblast Lineage Cells (Osterix+) is Required in Mice for Fracture Healing. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 34(9), 1690-1706.

+ Open protocol

+ Expand

Vessel Density Quantification in Tumor Sections

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tumor sections (5 μm), two sections per tumor, were stained with anti-PECAM-1 (M-20) (Santa Cruz Biotechnology). Bound antibody was detected with ImmPRESS (Peroxidase) Polymer anti-rabbit immunoglobulin reagent (Vector Laboratories) and visualized using ImmPACT DAB peroxidase (HRP) substrate (Vector Labs). Mayer's hematoxylin (Sigma-Aldrich) was used as a counterstain. Slides were digitized using an Aperio ScanScope XT scanner (Leica), and computer-aided image analysis was performed and manually checked for quality assurance. Regions of interest were identified with an algorithm that distinguishes tumor from stroma and (peri-)necrotic regions. The vessel density within the tumor region, as well as in the adjacent normal tissue, was quantified using the Definiens Tissue Studio software platform (Definiens). Quantification was done while blinded to the treatment groups.

Hammoud L., Adams J.R., Loch A.J., Marcellus R.C., Uehling D.E., Aman A., Fladd C., McKee T.D., Jo C.E., Al-Awar R., Egan S.E, & Rossant J. (2016). Identification of RSK and TTK as Modulators of Blood Vessel Morphogenesis Using an Embryonic Stem Cell-Based Vascular Differentiation Assay. Stem Cell Reports, 7(4), 787-801.

+ Open protocol

+ Expand

RNAscope and Immunohistochemistry for OA and RA

Check if the same lab product or an alternative is used in the 5 most similar protocols

RNAscope in situ hybridization was carried out on formalin-fixed paraffin-embedded human joint replacement tissues sections of OA and RA synovium provided by the Royal Orthopaedic Hospital (ethical approval no. 07/H1204/191). Sections were deparaffinized in xylene, followed by 100% ethanol. Detection of human C15orf48 transcript was performed using an RNAscope 2.5 HD manual assay red kit reagents and methods according to the manufacturer (ACD Bio-Techne). Following the last wash for the RNAscope protocol, slides were washed in distilled water for 5 min and then blocked with Bloxall (Vector Laboratories) for 10 min, followed by incubation with 10% normal horse serum in tris buffer for 10 min. Immunohistochemistry was performed by incubating sections overnight at 4°C with biotin-conjugated mouse anti-human CD68 (Novus, NBP2-34661B), followed by streptavidin-HRP (Thermo Fisher Scientific) and ImmPACT DAB Peroxidase (HRP) Substrate (Vector Laboratories). Hematoxylin QS (Vector Laboratories) was used for counterstain. Images were obtained using the Zeiss Axio Scan and analyzed in Zen Blue (both Zeiss).

Clayton S.A., Daley K.K., MacDonald L., Fernandez-Vizarra E., Bottegoni G., O’Neil J.D., Major T., Griffin D., Zhuang Q., Adewoye A.B., Woolcock K., Jones S.W., Goodyear C., Elmesmari A., Filer A., Tennant D.A., Alivernini S., Buckley C.D., Pitceathly R.D., Kurowska-Stolarska M, & Clark A.R. (2021). Inflammation causes remodeling of mitochondrial cytochrome c oxidase mediated by the bifunctional gene C15orf48. Science Advances, 7(50), eabl5182.

+ Open protocol

+ Expand

Immunohistochemical Identification of Chloride Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Chloride cells were targeted for immunohistochemical identification according to Adams and Marìn de Mateo (1994 ). Chloride cell staining used gill tissue fixed as previously described. Sections were de-waxed in xylene (10 min) and rehydrated through a graded ethanol series (100% for 5 min and 70% for 3 min). Sections were then blocked for endogenous peroxidase activity (3% H₂O₂ in methanol – 20 min), washed with PBS (5 min) and incubated with 10% normal goat serum (20 min) to block non-specific binding sites. Sections were blotted dry and incubated overnight in a humid chamber with a mouse monoclonal antibody to Na+/K+-ATPase (1:200, IgGα5, Developmental Studies Hybridoma Bank, Department of Biological Sciences, University of Iowa, Iowa, USA). Sections were washed with PBS and incubated (1 h, room temperature) with 1:200 goat anti-mouse IgG (A4416, Sigma, USA) before being washed again. ImmPACT DAB Peroxidase (HRP) Substrate (SK-4105; Vector Laboratories, Burlingame) was applied as per manufacturer instructions and sections were incubated for 5 min. Slides were then immersed in tap water (2–5 min) to stop the reaction, counterstained with haematoxylin Z (3 min), dehydrated, cleared and mounted.

CHALMERS L., TAYLOR J.F., ROY W., PRESTON A.C., MIGAUD H, & ADAMS A. (2017). A comparison of disease susceptibility and innate immune response between diploid and triploid Atlantic salmon (Salmo salar) siblings following experimental infection with Neoparamoeba perurans, causative agent of amoebic gill disease. Parasitology, 144(9), 1229-1242.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Tumor Protein Expression

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tumors were fixed in 4% paraformaldehyde overnight at 4°C. Then tumors were embedded in paraffin and sectioned at 5 μm. Immunohistochemistry was performed following standard procedures. After incubated with primary antibodies (Rabbit anti-GR, CST, #12041; mouse anti-NR2F1 R&D, PP-H8132-00), VECTASTAIN® ABC HRP Kit (Peroxidase, Rabbit IgG) and HRP conjugated Goat anti-mouse IgG were used, followed by ImmPACT® DAB Peroxidase (HRP) Substrate. Images were acquired on ECHO revolve microscope. Representative images of four tumors of each group were used to quantify the IHC signals of GR and NR2F1, using imageJ and the IHC Profiler plugin (Varghese et al., 2014 (link)).

Zhang Z., Zhou C., Li X., Barnes S.D., Deng S., Hoover E., Chen C.C., Lee Y.S., Zhang Y., Wang C., Metang L.A., Wu C., Tirado C.R., Johnson N.A., Wongvipat J., Navrazhina K., Cao Z., Choi D., Huang C.H., Linton E., Chen X., Liang Y., Mason C.E., de Stanchina E., Abida W., Lujambio A., Li S., Lowe S.W., Mendell J.T., Malladi V.S., Sawyers C.L, & Mu P. (2020). Loss of CHD1 Promotes Heterogeneous Mechanisms of Resistance to AR-Targeted Therapy via Chromatin Dysregulation. Cancer Cell, 37(4), 584-598.e11.

+ Open protocol

+ Expand

Immunohistochemical Analysis of PPARα in Liver

Check if the same lab product or an alternative is used in the 5 most similar protocols

Liver tissues were fixed in buffered 4% formalin for 48 h and then embedded in paraffin. Sections were deparaffinized and hydrated. Heat-mediated antigen retrieval was performed with 10 mM citrate buffer pH 6.0 (Thermo Fisher Scientific). Endogenous peroxide was inhibited by incubating with a freshly prepared 3% H₂O₂ solution in MeOH. Unspecific antigens were blocked by incubating sections for 1 h with 2.5% horse serum (Cat# VE-S-2000; Vector Laboratories, Burlingame, CA, USA). To assess the PPARα expression, 5 μm liver sections were stained with rabbit anti-mouse PPARα (1:400; Cat# ab61182; Abcam), followed by a goat anti-rabbit or HRP conjugate (ImmPRESS; Vector Laboratories). Color was developed after incubation with 3,3′-diaminobenzidine (DAB) substrate (Cat# SK-4105; Vector Laboratories; ImmPACT DAB Peroxidase (HRP) substrate), followed by hematoxylin counterstaining and mounting (Cat# Vecmount H-5000; Vector Laboratories). Stained sections were photographed as previously described. The positive area was quantified using ImageJ with a minimum of 5–6 random liver sections per mouse.

Azar S., Udi S., Drori A., Hadar R., Nemirovski A., Vemuri K.V., Miller M., Sherill-Rofe D., Arad Y., Gur-Wahnon D., Li X., Makriyannis A., Ben-Zvi D., Tabach Y., Ben-Dov I.Z, & Tam J. (2020). Reversal of diet-induced hepatic steatosis by peripheral CB1 receptor blockade in mice is p53/miRNA-22/SIRT1/PPARα dependent. Molecular Metabolism, 42, 101087.

+ Open protocol

+ Expand

Comprehensive Immunohistochemistry Protocol for Vascular Markers

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

IHC was performed as we previously described [47 (link)]. In brief, paraffin-embedded arteries were cut into 5-μm sections. Slides were deparaffinized and rehydrated through xylenes and graded alcohol series. Citrate buffer was used for antigen retrieval in a high-pressure cocker (2 h at 80 °C). Endogenous peroxidase was blocked via incubation with 3% H₂O₂ for 15 min. ImmPRESS™ HRP Anti-Rabbit IgG (Peroxidase) Polymer Detection Kit (Vector Laboratories, MP-7451-15) and primary antibodies were used for immunostaining of c-MYB, KLF4, αSMA, SM22, Calponin, and MYH11, which was visualized using ImmPACT DAB Peroxidase (HRP) Substrate. Six fields on each section were imaged.

Xie X., Shirasu T., Li J., Guo L.W, & Kent K.C. (2023). miR579-3p is an inhibitory modulator of neointimal hyperplasia and transcription factors c-MYB and KLF4. Cell Death Discovery, 9, 73.

+ Open protocol

+ Expand

Histological Characterization of Chondrocyte Pellets

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

After 21 days, pellets were washed with PBS and fixed in 10% neutral buffered formalin followed by paraffin embedding. Cross sections (5 μm thick) were stained with Safranin O-Fast Green for sulfated glycosaminoglycans (sGAG) using standard protocols. Immunohistochemical analysis was performed using a Collagen Type II (Col II) antibody (Developmental Studies Hybridoma Bank), followed by a goat anti-mouse-HRP (Jackson ImmunoResearch). Slides were developed using the ImmPACT DAB Peroxidase (HRP) Substrate (Vector Laboratories) and counterstained with VECTOR Hematoxylin QS (Vector Laboratories) per manufacturer’s instructions. Slides were visualized and images taken using a Nikon Eclipse 90i microscope (Tokyo, Japan). To measure total sGAG content, pellets were washed with PBS and each dry pellet was frozen at −80°C until ready to be assayed. sGAG content per pellet was quantified via the dimethylmethylene blue (DMMB) assay as previously described (n=3 pellets/group).^{11 (link)}

Aisenbrey E.A., Bilousova G., Payne K, & Bryant S.J. (2019). Dynamic mechanical loading and growth factors influence chondrogenesis of induced pluripotent mesenchymal progenitor cells in a cartilage-mimetic hydrogel. Biomaterials science, 7(12), 5388-5403.

+ Open protocol

+ Expand

Neutralizing Antibody Titration for LOM Virus

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Anti-CSF-neutralizing antibodies were detected by a neutralizing peroxidase-linked assay (NPLA) according to the standards manual of the World Organization for Animal Health (WOAH) [26 (link)]. Heat-inactivated serum samples (56 °C, 30 min) were serially diluted 2-fold (1:2 to 1:2048) and neutralized by adding 200 TCID₅₀ of the LOM virus for 1 h at 37 °C. The neutralized serum containing virus was inoculated into PK-15 cells (porcine kidney cells) and cultured for 3 days in a 37 °C incubator. The cells were then fixed in chilled 80% acetone and reacted with a commercial anti-LOM mAb (MEDIAN diagnostic Co., Cat No. 9013, Chuncheon, Republic of Korea). The cells were then stained using a VECTASTAIN^® ABC-HRP kit (Vector Laboratories Inc., Cat no. PK-4000, Newark, CA, USA) and an ImmPACT DAB Peroxidase (HRP) substrate (Vector Laboratories Inc., Cat no. PK-4100, USA). Neutralizing antibody titers were expressed as the reciprocal of the highest dilution that yielded 50% neutralization and determined to ≥10-fold positive antibodies according to the WOAH criteria.

Park G.N., Shin J., Choe S., Kim K.S., Kim J.J., Lim S.I., An B.H., Hyun B.H, & An D.J. (2023). Safety and Immunogenicity of Chimeric Pestivirus KD26_E2LOM in Piglets and Calves. Vaccines, 11(10), 1622.

+ Open protocol

+ Expand

Immunohistochemical Analysis of ASF1B

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tumor tissue sections were harvested and treated as previously described^{31 (link)}. Primary antibodies for anti-human ASF1B (1:100), anti-mouse ASF1B, Ki67, and CD31 (1:100) were purchased from Cell Signaling Technologies. An ImmPRESS™ HRP Anti-Rabbit IgG (Peroxidase) Polymer Detection Kit, raised in horse (Vector Laboratories), was used for the secondary antibody. ImmPACT DAB Peroxidase (HRP) Substrate was used for detection, and hematoxylin (Vector Laboratories) was used as a counterstain.

Liu X., Song J., Zhang Y., Wang H., Sun H., Feng X., Hou M., Chen G., Tang Q, & Ji M. (2020). ASF1B promotes cervical cancer progression through stabilization of CDK9. Cell Death & Disease, 11(8), 705.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!