Abc ap

Manufactured by Vector Laboratories

Sourced in United States

The ABC-AP is a laboratory instrument designed for the analysis of biological samples. It provides accurate and reliable measurements to support various research and testing applications. The core function of the ABC-AP is to perform quantitative analysis of target analytes in a controlled and standardized manner.

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Lab products found in correlation

Ak 5000, by Vector Laboratories (1 mentions) Ba 9200, by Vector Laboratories (1 mentions) Vector red substrate, by Vector Laboratories (1 mentions) Biotinylated secondary antibody, by Vector Laboratories (1 mentions) P nitrophenyl phosphate substrate, by Merck Group (1 mentions) M3 plate reader, by Molecular Devices (1 mentions) Target retrieval system, by Agilent Technologies (1 mentions) Vectastain abc ap kit, by Vector Laboratories (1 mentions) Liquid permanent red, by Agilent Technologies (1 mentions)

Close spelling variants of this product

Vectastain ABC-AP alkaline phosphatase Vectastain ABC-AP standard kit Vectastain ABC-AP kit Vectastain ABC-AP staining-kit Vectastain ABC-AP Rabbit IgG Kit Vectastain Universal ABC-AP kit ABC-AP Mouse IgG kit ABC-AP reagent ABC-AP) (AK-5000), ABC-AP kit

8 protocols using abc ap

Immunodetection of T. bryosalmonae

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Tissue sections of anterior and posterior kidneys, liver, and spleen of all 31 brown trout underwent immunohistochemistry to detect T. bryosalmonae antigens. Heat-induced epitope retrieval (microwave pressure cooker for 33 min) with a tris buffer at pH 9.0 was used for antigen demasking, followed by avidin biotin blocking and normal goat serum to block unspecific reactions. After determination of the final dilution, a monoclonal mouse antibody (IgG1, P01, Aquatic Diagnostics Ltd, Scotland) was used as the primary antibody (1:50 in tris-buffered saline). To increase sensitivity a biotinylated goat anti-mouse antibody served as the secondary antibody (Vector, BA-9200, Burlingame, CA, USA). After incubation with avidin-coupled alkaline phosphatase (ABC-AP, Vector, AK-5000, Burlingame, CA, USA), Liquid Permanent Red (K0640, Dako Agilent, Glostrup, Denmark) was used as the chromogen. Positive (PKD-infected fish) and negative (substitution of the primary antibody with an irrelevant mouse IgG) controls were included in each assay.

Arndt D., Fux R., Blutke A., Schwaiger J., El-Matbouli M., Sutter G, & Langenmayer M.C. (2019). Proliferative Kidney Disease and Proliferative Darkening Syndrome are Linked with Brown Trout (Salmo trutta fario) Mortalities in the Pre-Alpine Isar River. Pathogens, 8(4), 177.

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Immunohistochemical Analysis of ER Stress Chaperons

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Hearts were excised from mice, embedded in paraffin, sectioned into 4 μm slices, and stained with H&E^{56 (link)}. To determine the expression level of ER stress chaperons, immunohistochemical staining was performed on paraffin-embedded sections with a primary antibody against the KDEL receptor (1:50 dilution), and then with a biotinylated secondary antibody (undiluted goat anti-rabbit/mouse (H+L), Dako Denmark A/S, Hovedstaden, Denmark)^{56 (link)}. The sections were then treated with peroxidase-conjugated biotin–avid in complex using VECTASTAIN ABC-AP and visualized by DAB (3, 3-diaminobenzidine).

Yao Y., Lu Q., Hu Z., Yu Y., Chen Q, & Wang Q.K. (2017). A non-canonical pathway regulates ER stress signaling and blocks ER stress-induced apoptosis and heart failure. Nature Communications, 8, 133.

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Immunohistochemical Analysis of AGGF1 and CD31 in Mouse Hearts

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Four weeks after AGGF1 or IgG treatment, mice were anesthetized, euthanized, and the hearts were excised and fixed overnight. The fixed hearts were sectioned and immunohistochemical staining was performed on paraffin-embedded sections with a primary antibody against AGGF1 or CD31, which was followed by incubation with a biotinylated secondary antibody as described previously [29 (link),37 (link)]. The sections were then treated with peroxidase-conjugated biotin–avidin complex using VECTASTAIN ABC-AP and visualized by DAB. Slides were counterstained with H&E staining. Vessel density was evaluated by counting the number of neovessels and arterioles in five random and non-repeated high power fields.

Lu Q., Yao Y., Hu Z., Hu C., Song Q., Ye J., Xu C., Wang A.Z., Chen Q, & Wang Q.K. (2016). Angiogenic Factor AGGF1 Activates Autophagy with an Essential Role in Therapeutic Angiogenesis for Heart Disease. PLoS Biology, 14(8), e1002529.

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Dual Immunocytochemistry for Megakaryocyte and Erythroid Lineages

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After 12–14 days, fixed and dried cultures were stained using a dual immunocytochemistry assay as previously described^{3 (link)}. E staining was completed using rabbit anti-GlyA antibody (1:1,500, ABD Serotec, clone YTH89.1), the horse anti-rabbit Impress-horseradish peroxidase secondary kit (Vector Labs) for signal amplification, and nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate. Mk staining was completed using mouse anti-human CD41a antibody (BD), goat anti-mouse Impress-AP secondary (Vector Labs), ABC-AP, and Vector Red substrate (Vector Labs). Colonies were scored based on GlyA and CD41a staining as megakaryocyte (Mk) only, erythroid (E) only, or megakaryocyte/erythroid (Mk/E).

Scanlon V.M., Thompson E.N., Lawton B.R., Kochugaeva M., Ta K., Mayday M.Y., Xavier-Ferrucio J., Kang E., Eskow N.M., Lu Y.C., Kwon N., Laumas A., Cenci M., Lawrence K., Barden K., Larsuel S.T., Reed F.E., Peña-Carmona G., Ubbelohde A., Lee J.P., Boobalan S., Oppong Y., Anderson R., Maynard C., Sahirul K., Lajeune C., Ivathraya V., Addy T., Sanchez P., Holbrook C., Van Ho A.T., Duncan J.S., Blau H.M., Levchenko A, & Krause D.S. (2022). Multiparameter analysis of timelapse imaging reveals kinetics of megakaryocytic erythroid progenitor clonal expansion and differentiation. Scientific Reports, 12, 16218.

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CD4-Positive Cell Immunohistochemistry in Ear Tissue

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Ear tissue sections were deparaffinized using xylene. Then, the sections were rehydrated with graded ethanol solutions and rinsed with deionized H₂O. Antigen retrieval was produced with Tris-EDTA buffer (10 mM Tris Base, 1 mM EDTA solution, 0.05% Tween 20, pH 9.0) using the microwave for 15 min. To eliminate endogenous peroxidase activity, the sections were treated with 3% hydrogen peroxide for 10 min. The sections were blocked with blocking buffer (5% normal goat serum in PBS with 0.05% Tween 20) for 1 h at room temperature and then incubated with primary antibody against CD4 (1:500, Abcam, Cambridge, UK) overnight at 4℃. Next, biotinylated secondary antibody (1:200, Vector Laboratories, Newark, CA, USA) was applied for 30 min, and ABC-AP (Vector Laboratories) reagent was applied for 30 min. Subsequently, the sections were incubated with AP substrate for 20 min and counterstained with hematoxylin for 1 min at room temperature. IHC-stained sections were observed under ×400 magnification.

Bae S., Jeong N.H., Choi Y.A., Lee B., Jang Y.H., Lee S, & Kim S.H. (2023). Lupeol alleviates atopic dermatitis-like skin inflammation in 2,4-dinitrochlorobenzene/Dermatophagoides farinae extract-induced mice. BMC Pharmacology & Toxicology, 24, 27.

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CL-K1 Plasma Quantification by ELISA

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Plasma CL-K1 was assayed using a previously established sandwich ELISA method using two CL-K1 specific antibodies, with minor modifications [6 (link)]. Briefly, 384 well plates were coated with 20 μl of an anti-CL-K1 rabbit polyclonal antibody. After rinsing and blocking, 20 μl of diluted plasma samples and standard were incubated in duplicate. All plasma samples used in this study were thawed only once. After rinsing, the wells were incubated with a biotinylated anti-CL-K1 monoclonal antibody followed by ABC-AP (Vector Lab, Burlingame, CA) and then developed using p-nitrophenyl phosphate substrate (Sigma-Aldrich). Reactions were assayed for absorbance at 405 nm using an M3 plate reader (Molecular Devices).
From assays performed using plasma specimens from 27 healthy volunteers, the median of plasma CL-K1 levels was 245 ng/ml with interquartile range 160–273 ng/ml. The highest concentration measured in this group was 772 ng/ml. The CL-K1 reference range was found to be <619 ng/ml (normal donor mean + 2SD).
Plasma samples from a subpopulation of 216 patients (201 DIC and 15 non-DIC) were also assayed for plasma CRP, an acute phase protein, using ELISA (DuoSet, R&D Systems), according to the manufacturer’s instructions.

Takahashi K., Ohtani K., Larvie M., Moyo P., Chigweshe L., Van Cott E.M, & Wakamiya N. (2014). Elevated plasma CL-K1 level is associated with a risk of developing disseminated intravascular coagulation (DIC). Journal of thrombosis and thrombolysis, 38(3), 331-338.

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Immunohistochemical Detection of TALDO1

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After deparaffinizing sections, antigens were retrieved using the Target Retrieval System (Dako, Inc.). Sections were blocked with horse serum and incubated with rabbit polyclonal anti‐TALDO1 (1:50; Abcam) or rabbit IgG control overnight at 4℃, with biotinylated anti‐rabbit IgG (Vectastain^®ABC‐AP Kit; Vector Labs), and then with ABC‐AP (Vector Labs). Slides were incubated with levamizole added to blue substrate working solution. Sections were imaged with a light microscope equipped with the Cri‐Nuance system (Perkin Elmer Corp, Inc.) to subtract out melanin pigment.

Cano M., Datta S., Wang L., Liu T., Flores‐Bellver M., Sachdeva M., Sinha D, & Handa J.T. (2021). Nrf2 deficiency decreases NADPH from impaired IDH shuttle and pentose phosphate pathway in retinal pigmented epithelial cells to magnify oxidative stress‐induced mitochondrial dysfunction. Aging Cell, 20(8), e13444.

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Immunohistochemical Detection of S100A4

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IHC was performed with polyclonal rabbit anti-S100A4 (Dako) on 4 μm xenograft tumor sections. Rabbit negative control (Dako) served as isotype control. Briefly, sections were dewaxed and antigen retrieval was carried out in citrate buffer (pH 6) at 60°C overnight. Non-specific binding was blocked by incubating the sections in 10% normal swine serum followed by incubation with the primary antibody (1:10 in antibody diluent, Dako). After washing, sections were incubated with biotinylated swine-anti-rabbit antibody. Detection of S100A4 antibody was performed by the streptavidin-alkaline phosphatase kit (ABC-AP; Vector Labs.). Enzyme reactivity of the AP complex was visualized using DAKO liquid permanent red.

Grottke A., Ewald F., Lange T., Nörz D., Herzberger C., Bach J., Grabinski N., Gräser L., Höppner F., Nashan B., Schumacher U, & Jücker M. (2016). Downregulation of AKT3 Increases Migration and Metastasis in Triple Negative Breast Cancer Cells by Upregulating S100A4. PLoS ONE, 11(1), e0146370.

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