Histofine simple stain rat max po multi

Manufactured by Nichirei Biosciences

Sourced in Japan

The Histofine Simple Stain Rat MAX PO (MULTI) is a laboratory equipment product designed for immunohistochemical staining procedures. It is a polymer-based detection system that enhances the visualization of target antigens in rat tissue samples.

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Lab products found in correlation

3 3 diaminobenzidine dab substrate kit, by Nichirei Biosciences (1 mentions) Axiophot 2, by Zeiss (1 mentions) Hydrogen peroxide, by Fujifilm (1 mentions) Nitrilotriacetic acid, by Fujifilm (1 mentions) Normal goat serum (ngs), by Vector Laboratories (1 mentions) 2 thiobarbituric acid tba, by Merck Group (1 mentions) Tween 20, by Merck Group (1 mentions) Trichloroacetic acid, by Merck Group (1 mentions) Glucose 6 phosphate (g6p), by Merck Group (1 mentions) Histosec pastilles, by Merck Group (1 mentions) Von kossa method for calcium kit, by Polysciences (1 mentions) Dab substrate kit, by Nichirei Biosciences (1 mentions) Ab39012, by Abcam (1 mentions) Liquid diaminobenzidine substrate chromogen system, by Agilent Technologies (1 mentions) Ab6992, by Abcam (1 mentions) M0851, by Agilent Technologies (1 mentions) Sc 166182, by Santa Cruz Biotechnology (1 mentions) Ab9722, by Abcam (1 mentions) Anti s100 polyclonal antibody, by Agilent Technologies (1 mentions) Antidesmin mouse monoclonal antibody clone d33, by Agilent Technologies (1 mentions)

Close spelling variants of this product

Histofine Simple Stain MAX PO (154 citations) Histofine Simple Stain MAX-PO MULTI (37 citations) Histofine Simple Stain (21 citations) Histofine Simple Stain Rat MAX PO (14 citations) Simple stain rat MAX-PO (10 citations) Simple stain MAX PO MULTI (7 citations) Histofine® MAX PO Multi (3 citations)

9 protocols using histofine simple stain rat max po multi

Immunostaining of Bombyx mori Vasa-like Gene

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Immunostainings were also performed by the New Histo. Science Laboratory Co. (Ome, Japan). After deparaffinization, slices were transferred into 10 mM citrate buffer solution (pH6.0) and incubated at 95°C for 20 min. After cooling down to room temperature, the slices were washed with PBS (pH 7.4) for 5 min. Then the slices were transferred into 3% hydrogen peroxide solution for 5 min at room temperature; washed three times in PBS (pH 7.4) for 5 min each followed by incubation with anti-BmVLG (B. mori vasa-like gene) antibody at a 1:100 dilution for 50 min at room temperature. After three-times washing with PBS (pH 7.4) for 5 min each, they were incubated with secondary antibody Histofine Simple Stain Rat MAX PO (MULTI) (NICHIREI BIOSCIENCES INC.) for 30 min at room temperature. After three-times washing with PBS (pH 7.4) for 5 min each, the signals were detected by incubation with 3,3′-Diaminobenzidine, tetrahydrochloride. After washing with diluted water, the nucleus was stained with Mayer hematoxylin solution for 1 minute.
Anti-BmVLG (BmVasa571) used in the present study was produced by immunizing rabbits with a synthetic peptide, CKGNKYGGSDVRNFN (Genscript).

Sakai H., Kirino Y., Katsuma S., Aoki F, & Suzuki M.G. (2016). Morphological and histomorphological structures of testes and ovaries in early developmental stages of the silkworm, Bombyx mori. Journal of insect biotechnology and sericology, 85(1), 15-20.

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Visualization of Collagen Proteins

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Proteins in the collagen sheet were visualized with antibodies of BMP-2 and osteopontin. The sections were washed in 10 nmol/L with pH 7.4 phosphate-buffered saline (PBS) and endogenous peroxidase activity was blocked by incubating sections with 0.3% H₂O₂ in methanol for 30 min.
The sections were then reacted with the primary antibodies, BMP-2 polyclonal antibody diluted 1:50 (Proteintech Group, Chicago, USA) and Anti-Osteopontin (rabbit) polyclonal antibody (R&D Systems, Minnesota, USA) diluted 1:100, by incubating at 37 °C for 60 min. The sections were washed in PBS and then incubated with the secondary antibody, peroxidase-labeled anti-mouse IgG polyclonal antibody (Histofine Simple Stain Rat MAX-PO [MULTI]; Nichirei, Tokyo, Japan) for 30 min and washed with PBS. Subsequently, the sections were stained with 3,3′-diaminobenzidine (DAB substrate kit, Nichirei, Tokyo, Japan), washed in sterilized water, and counterstained with hematoxylin. The sections were then dehydrated according to established protocol and the sections were examined and photographed using a universal photomicroscope (Axiophot 2).

Kanda Y., Nishimura I., Sato T., Katayama A., Arano T., Ikada Y, & Yoshinari M. (2016). Dynamic cultivation with radial flow bioreactor enhances proliferation or differentiation of rat bone marrow cells by fibroblast growth factor or osteogenic differentiation factor. Regenerative Therapy, 5, 17-24.

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Immunohistochemical Analysis of CD90 Expression

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The streptavidin–biotin immunoperoxidase method was employed for immunohistochemistry using the Histofine simple stain rat MAX‐PO (MULTI; Nichirei Bioscience Co., Ltd., Tokyo, Japan). Sections were deparaffinised with xylene, washed with graded ethanol, and then washed again with distilled water. Sections were microwaved at 65°C for 20 min in 0.01 M citrate buffer (pH 6.0), cooled to room temperature (RT) and washed in distilled water. Endogenous peroxidase activity was blocked by incubating the sections with 0.3% H₂O₂ in methanol for 30 min and then washing in phosphate‐buffered saline (PBS, pH7.2, 0.01M) three times for 5 min each. In order to prevent non‐specific reactions, sections were incubated with 10% goat serum for 30 min. Anti‐rat CD90 (Thy‐1; dilution 1:100, eBioscience, CA, USA) mouse antibody was incubated at RT for 1 h. Horseradish peroxidase (HRP)‐conjugated anti‐mouse IgG was then incubated at RT for 30 min. After washing in PBS three times for 5 min each, 3,3′‐diaminobenzidine‐tetrahydrochloride (DAB) in 0.05 M Tris–HCl buffer (pH7.6) was used to visualise reactivity. After washing in distilled water, sections were counter‐stained with haematoxylin for 30 s and then observed under a light microscope (Axiophot2; Carl Zeiss, Oberkochen, Germany). Drilled dentine of the upper right first molar and the observation area were shown in Fig. 1.

Sano Y., Sugiuchi A., Mitomo K., Yanagisawa A., Kambe R., Furusawa M, & Muramatsu T. (2018). Changes of CD90 expression and immunoreactive cell localisation in rat dental pulp after cavity preparation. Australian Endodontic Journal, 45(2), 189-195.

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Antioxidant Enzyme Activity Assay

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Iron nitrate enneahydrate, nitrilotriacetic acid, oxidized and reduced glutathione, glutathione reductase, and hydrogen peroxide were purchased from Wako (Osaka, Japan). Nicotinamide adenine dinucleotide phosphate reduced tetrasodium salt, glucose-6-phosphate, 1-chloro-2,4-dinitrobenzene (CDNB), 5,5'-dithio-bis-2-nitrobenzoic acid (DTNB), bovine serum albumin, trichloroacetic acid, and Tween 20 were purchased from Sigma (St. Louis, MO). 2-Thiobarbituric acid (TBA) was purchased from Merck (Darmstadt, Germany). The basal MF diet (powder) and Vitamin E (VE)-deficient diet were purchased from Oriental Yeast (Tokyo, Japan). The VE-stripped corn oil was purchased from TAMA Biochemicals (Tokyo, Japan). The BCA assay kit was purchased from Pierce (Rockford, IL). The Ax-C-8 was a kind gift from the Institute for Health Care Science, Suntory (Osaka, Japan). Normal goat serum was purchased from Vector Laboratories (Burlingame, CA), and Histofine Simple Stain rat Max-PO (multi) was purchased from Nichirei Biosciences (Tokyo, Japan). Liquid DAB was purchased from DAKO Japan (Tokyo, Japan). The two monoclonal antibodies against 8-OHdG (N45.1) and HNE-modified proteins (HNE-J2) were from the Japan Institute for the Control of Aging (Shizuoka, Japan). All of the other chemicals were of the highest quality available from Wako (Osaka, Japan).

Okazaki Y., Okada S, & Toyokuni S. (2017). Astaxanthin ameliorates ferric nitrilotriacetate-induced renal oxidative injury in rats. Journal of Clinical Biochemistry and Nutrition, 61(1), 18-24.

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Immunohistochemical Detection of eIF-2α

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Samples were fixed for 16 hr at 4°C with 4% paraformaldehyde in 0.1 M sodium phosphate buffer (pH 7.4), washed with the same buffer, dehydrated in a graded series of ethanol concentrations followed by xylene, then embedded in paraffin wax (Histosec^® pastilles, Merck KGaA, Darmstadt, Germany). Thin sections (5 μm) were cut using a microtome. For detection of eIF-2α, deparaffinized sections were immersed in 10 mM citrate buffer and microwaved for 20 min at 800 W (MicroMed T/T; Milestone, Sorisole, Italy). After antigen retrieval, the sections were blocked with 1% BSA-PBS for 1 hr at 20–22°C. These sections were then incubated for 16 hr at 4°C with anti-eIF-2α and anti-phospho-eIF-2α antibodies diluted 1:100 in 1% BSA-PBS. After three washes with PBS, the indirect immunoperoxidase method (Histofine Simple Stain Rat MAX-PO-Multi; Nichirei Bioscience) was applied to the sections. The sections were then treated for 7 min with a freshly prepared solution of 0.1 mg/ml 3, 3′-diaminobenzidine (DAB) in 50 mM Tris-HCl buffer (pH 7.6) containing 0.05% hydrogen peroxide to visualize eIF-2α and further stained with hematoxylin. The specificity of immunoreactivity was confirmed by replacing the primary antibodies with 1% BSA-PBS.

Nakamata J., Morimoto H., Baba R., Kokubu K, & Miyamoto T. (2024). Glucose Induces ER Stress Response-Mediated Peritoneal Mesothelial Cell Death. Acta Histochemica et Cytochemica, 57(1), 7-14.

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Thoracoabdominal Aorta Calcification Analysis

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Approximately one third of the collected thoracoabdominal aorta was used to visualize calcification and conduct immunostaining. The aorta was embedded in paraffin and 4-µm sections were cut. Sections were stained using a Von Kossa Method for Calcium Kit (Polysciences, Warrington, PA) according to the manufacturer’s instructions. Immunostaining analysis was performed with serial sections. The sections were incubated with SLC37A2 antibody overnight at 4°C and then treated with Histofine Simple Stain Rat MAX-PO (MULTI) (Nichirei Biosciences, Tokyo, Japan) for 30 min at room temperature. Thereafter, sections were treated with a DAB substrate kit (Nichirei).
Total RNA was extracted from the rest of the frozen thoracoabdominal aorta. Real-time PCR was performed using the synthesized cDNA. The primer sequences were as follows: rat osteopontin, 5'-ACCCATCTCAGAAGCAGAATC-3' and 5'-ATC CATGTGGTCATGGCTTTC-3' and; rat SLC37A2, 5'-TAG TCCCAGCTTCGAGTACGG-3' and 5'-CTGATGGGCTTTCTG GACATG-3'. The osteopontin mRNA and SLC37A2 mRNA expression levels were calculated after normalization with GAPDH.

Tani M., Tanaka S., Oeda C., Azumi Y., Kawamura H., Sakaue M, & Ito M. (2020). SLC37A2, a phosphorus-related molecule, increases in smooth muscle cells in the calcified aorta. Journal of Clinical Biochemistry and Nutrition, 68(1), 23-31.

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Immunohistochemistry Analysis of Rib Fracture

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For immunohistochemistry, dehydrated sections of rib fracture were treated with 0.3% H₂O₂ in phosphate-buffered saline (PBS; pH 7.4) for 30 min at room temperature to inactivate endogenous peroxidase. Sections were pretreated with 3% bovine serum albumin (BSA) in PBS for 30 min at room temperature, followed by incubation with primary antibodies against IL-1β (1:100, ab9722, Abcam, Cambridge, MA, USA), RARγ (1:100, #8965S, Cell Signaling Technology, Danvers, MA, USA), MMP-13 (1:100, ab39012, Abcam), CCN2 (1:100, ab6992, Abcam), α-SMA (1:1000, M0851, DakoCytomation, Glostrup, Denmark), and p-p38 (1:100, sc166182, Santa Cruz Biotechnology, Santa Cruz, CA, USA) overnight at 4 °C. Sections were reacted with Histofine Simple Stain rat MAX-PO (Multi; Nichirei, Tokyo) for 1 h at room temperature. Color was developed with the use of a liquid diaminobenzidine substrate-chromogen system (Dako, Carpinteria, CA, USA). Immunostained sections were then counterstained with methylene green.

Shimo T., Takebe H., Okui T., Kunisada Y., Ibaragi S., Obata K., Kurio N., Shamsoon K., Fujii S., Hosoya A., Irie K., Sasaki A, & Iwamoto M. (2020). Expression and Role of IL-1β Signaling in Chondrocytes Associated with Retinoid Signaling during Fracture Healing. International Journal of Molecular Sciences, 21(7), 2365.

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Immunohistochemistry Panel for Pleural Mesothelioma

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Anti-podoplanin rabbit polyclonal antibody (KS-17) was from Sigma (St. Louis, MO). Anti-multi-cytokeratin mouse monoclonal antibody (RTU-AE1/AE3) was from Novocastra (Newcastle, UK). Anti-desmin mouse monoclonal antibody (clone D33) was from DAKO (Carpinteria, CA). Anti-calretinin rabbit monoclonal antibody (clone SP13) was from Abcam (Cambridge, MA). Anti-S-100 polyclonal antibody was from DAKO. Anti-C-ERC/mesothelin monoclonal antibody was from IBL (Takasaki, Gunma, Japan). Anti-WT1 rabbit polyclonal antibody (C-19) was from Santa Cruz (Santa Cruz, CA). Secondary antibody was Histofine Simplestain rat MAX-PO (Multi) from Nichirei Bioscience (Tokyo).

AIERKEN D., OKAZAKI Y., HWU CHEW S., SAKAI A., WANG Y., NAGAI H., MISAWA N., KOHYAMA N, & TOYOKUNI S. (2014). RAT MODEL DEMONSTRATES A HIGH RISK OF TREMOLITE BUT A LOW RISK OF ANTHOPHYLLITE FOR MESOTHELIAL CARCINOGENESIS. Nagoya Journal of Medical Science, 76(1-2), 149-160.

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Immunohistochemical Detection of ANXA2

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Sections that were cut at a 4-mm thickness were dewaxed with xylene and ethanol, and subjected to high-temperature antigen retrieval in an immunosaver (Nisshin EM, Tokyo, Japan) for 40 min. Next, the slides were dipped for 30 min in methanol containing H 2 O 2 (0.3% v/v) to quench endogenous peroxidase activity. After washing with PBS, the slides were incubated with primary antibodies and washed with PBS three times for 5 min each, and Histofine Simple Stain MAX-PO (Multi) (Nichirei, Tokyo, Japan) or Histofine Simple Stain Rat MAX-PO (Multi) (Nichirei) was applied to the slides. Anti-ANXA2 rabbit polyclonal antibody (ab41803) was used at a final concentration of 0.5 mg/ml. Localization of antigen-antibody complexes was visualized as brown precipitates using DAB þ Liquid (Dako, Kyoto, Japan) before nuclear counterstaining with hematoxylin.

Yamashita K., Nagai H, & Toyokuni S. (2015). Receptor role of the annexin A2 in the mesothelial endocytosis of crocidolite fibers. Laboratory investigation; a journal of technical methods and pathology, 95(7).

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