Sk 4800

Manufactured by Vector Laboratories

Sourced in Canada

The SK-4800 is a precision pipette-based protein transfer device designed for efficient and consistent protein sample delivery. It features an adjustable volume range and is suitable for a variety of laboratory applications.

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Lab products found in correlation

Phosphor chk2, by Cell Signaling Technology (1 mentions) Gh2ax, by Cell Signaling Technology (1 mentions) Mkb 2225, by Vector Laboratories (1 mentions) Ba 1000, by Vector Laboratories (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Pk 6100, by Vector Laboratories (1 mentions) Hematoxylin, by Thermo Fisher Scientific (1 mentions) Monoclonal mouse anti p53, by Abcam (1 mentions) Cytoseal, by Thermo Fisher Scientific (1 mentions) Anti goat igg, by Vector Laboratories (1 mentions) Pk 7800, by Vector Laboratories (1 mentions) Anti tryptophan hydroxylase immunohistochemistry t8575, by Merck Group (1 mentions) Vectra, by PerkinElmer (1 mentions) Vectastain, by Vector Laboratories (1 mentions) Fluoromount g, by Southern Biotech (1 mentions) Ab31630, by Abcam (1 mentions) Ab182422, by Abcam (1 mentions) Axio imager z2, by Zeiss (1 mentions) Ab152, by Merck Group (1 mentions) K4003, by Agilent Technologies (1 mentions)

7 protocols using sk 4800

Immunofluorescence and Immunohistochemistry Protocols

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Immunofluorescence was performed based on manufacturer’s instructions and as per previously published protocols (74 (link)). Antibodies used include phosphor-Chk2 (Cell Signaling Technology, catalog no. 2197S), ER (EMD Millipore, catalog no. 04820MI), 53BP1 (Novus Biologicals, catalog no. NB100-304), gH2AX (Cell Signaling Technology, catalog no. 9718), and Ki67 (Abcam, catalog no. ab16667). Immunohistochemistry was performed on the basis of manufacturer’s instructions. Sections were first deparaffinized, then endogenous peroxidases were quenched using 3% H₂O₂, and antigen retrieval was done using 1× citric acid buffer. The blocking buffer used was 2% goat serum. Antibodies used were ER, Ki67 (Abcam, catalog no. ab16667), cleaved caspase-3 (Cell Signaling Technology, catalog no. 9664), and PR (Thermo Fisher Scientific, catalog no. MA512658). Primary antibodies were left overnight in 4° and followed by anti-rabbit secondary (Vector Laboratories, catalog no. BA-1000) or anti-mouse secondary (Vector Laboratories, catalog no. MKB-2225). Next, sections were incubated in avidin-biotin complex solution (Vector Laboratories, catalog no. PK-6100), stained with peroxidase substrate (Vector Laboratories, catalog no. SK-4800), and counterstained in hematoxylin. Images were captured on an Echo Revolve microscope.

Oropeza E., Seker S., Carrel S., Mazumder A., Lozano D., Jimenez A., VandenHeuvel S.N., Noltensmeyer D.A., Punturi N.B., Lei J.T., Lim B., Waltz S.E., Raghavan S.A., Bainbridge M.N, & Haricharan S. (2023). Molecular portraits of cell cycle checkpoint kinases in cancer evolution, progression, and treatment responsiveness. Science Advances, 9(26), eadf2860.

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Immunohistochemistry for Protein Localization

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IHC was also used for localization of biomarkers and to detect differentially expressed proteins when autofluorescence and light scattering interfered with image quality. IHC was used for DLX5, DLX6, HAND2 and P53. For IHC, sections were deparaffinized and rehydrated in a series of ethanol dilutions. The sections were boiled for 30 seconds in 10% blocking solution (Biocare, MX, DV-200), washed with 0.05% PBS-Triton, then incubated with the following primary antibodies: rabbit polyclonal anti-DLX5 (Abcam, MA, YH05211C), polyclonal goat anti-DLX6 (Santa Cruz, CA, sc-18154), monoclonal mouse anti-HAND2 (Santa Cruz, CA, sc-130629), monoclonal mouse anti-P53 (Abcam, MA, GR1277-1), diluted in horse serum overnight at 4 °C. Next day, they were washed with 0.05% PBS-Triton and then incubated with the secondary anti-mouse IgG, anti-rabbit IgG, or anti-goat IgG (Vector lab PK-7800, CA) for 20 minutes at room temperature and then washed with 0.05% PBS-Triton. We then treated the samples with tertiary antibody streptavidin-peroxidase complex (Vector lab SK-4800, CA) for 5 minutes at room temperature. Finally, the sections were treated with chromagen (Impact Novared Kit SK-4805) followed with hematoxylin (Fisher scientific, 123022) for nuclear staining. The hematoxylin was rinsed off with distilled water and the slides mounted with cytoseal (Fisher Scientific, PA).

Fakhouri W.D., Metwalli K., Naji A., Bakhiet S., Quispe-Salcedo A., Nitschke L., Kousa Y.A, & Schutte B.C. (2017). Intercellular Genetic Interaction Between Irf6 and Twist1 during Craniofacial Development. Scientific Reports, 7, 7129.

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Histochemical Mapping of Serotonin Neurons

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Rats were deeply anesthetized using isoflurane and final electrode coordinates were marked by passing current from a 6 V battery through 4 of the 16 nichrome electrode wires. Rats were perfused with 0.9% biological saline and 4% paraformaldehyde in a 0.2 M Potassium Phosphate Buffered Solution. Brains were extracted and post-fixed in a 10% neutral-buffered formalin solution for 24 hr, stored in 10% sucrose/formalin and sectioned via microtome. All brains processed for light microscopy using anti-tryptophan hydroxylase immunohistochemistry (T8575, Sigma-Aldrich, St. Louis, MO) and a NovaRed chromagen reaction (SK-4800, Vector Laboratories, Burlingame, CA) Sections were mounted, imaged using a light microscope and electrode placement was confirmed (Paxinos and Watson, 2007 ).

Wright K.M, & McDannald M.A. (2019). Ventrolateral periaqueductal gray neurons prioritize threat probability over fear output. eLife, 8, e45013.

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Histological Analysis of Mouse Colon

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Mouse colon was isolated, washed in PBS and dehydrated in ethanol series, xylene and paraffin. Afterwards, the tissue was embedded in paraffin, formalin fixed, and paraffin embedded (FFPE) and sections of 5 μm were cut. Haematoxylin and eosin (HE) and Masson's trichrome (MT) staining were performed using standard techniques. For the immunohistochemistry, the tissue was heated in citrate buffer (pH 6.0, 1.0 nmol/L citric acid) and endogenous peroxidase activity was quenched with 3% H₂O₂ in methanol. Standard indirect immunoperoxidase procedures were used for detection (Vectastain and SK-4800, Vector Laboratories) with the antibody Rabbit-αKi-67 (1:100, Novus Biologicals), Rat-αF4/80 (1:500, Caltag laboratories and slides were counterstained with Mayer's haematoxylin. Followed by mounting using Quick D mounting medium. For fluorescent staining, slides were incubated with Rabbit-αCASPIII antibody (1:100, ab2302, Abcam) and nuclear counterstain with DAPI using Fluoromount-G (0100-01, Southern Biotech). Slides were imaged using the PerkinElmer Vectra (Vectra 3.0.3; PerkinElmer, MA) at 20x magnification or imaged using the Leica SP8 with a water dipping objective.

Muntjewerff E.M., Tang K., Lutter L., Christoffersson G., Nicolasen M.J., Gao H., Katkar G.D., Das S., ter Beest M., Ying W., Ghosh P., Aidy S.E., Oldenburg B., van den Bogaart G, & Mahata S.K. (2021). Chromogranin A regulates gut permeability via the antagonistic actions of its proteolytic peptides. Acta physiologica (Oxford, England), 232(2), e13655.

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Quantitative Histological Analysis of Kidney Graft Macrophages

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Kidney remnants and explanted grafts were fixed in 4% PFA and embedded into paraffin for sectioning (3 μm thickness) before staining. Hematoxylin and Eosin, Masson's trichrome Verhoeff, Periodic Acid Schiff (PAS), von Kossa and Picro Sirius Red (SR) stainings were performed using standard techniques. Graft sections were stained (60 min at RT) using CD68 (pan-macrophage marker, ab31630, 1:250; Abcam,) and CD163 (M2 macrophage marker; ab182422, 1:500; Abcam) and visualized with NovaRED (SK-4800, Vector Laboratories). Whole cross sections. were scanned and digitized with the AxioScan M200682 scanner (Carl Zeiss Microimaging Inc.) and analyzed by two blinded scorers with ImageJ open software. The positive surface area was measured by selecting the whole tissue cross-section as area of interest, separating the area stained with NovaRED by using the color deconvolution plugin and setting a threshold. We then calculated the % positive surface area by dividing the separated NovaRED positive area by the total tissue area ×100. Cell nuclei were separated by water shedding and subsequently counted using ImageJ open software.

Besseling P.J., Krebber M.M., Fledderus J.O., Teraa M., den Ouden K., van de Kaa M., de Bree P.M., Serrero A., Bouten C.V., Dankers P.Y., Cox M.A, & Verhaar M.C. (2023). The effect of chronic kidney disease on tissue formation of in situ tissue-engineered vascular grafts. APL Bioengineering, 7(2), 026107.

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Immunohistochemical Confirmation of Electrode Placement

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Rats were deeply anesthetized using isoflurane and final electrode coordinates were marked by passing current from a 6V battery through 4 of the 16 nichrome electrode wires. Rats were perfused with 0.9% biological saline and 4% paraformaldehyde in a 0.2 M Potassium Phosphate Buffered Solution. Brains were extracted and postfixed in a 10% neutral-buffered formalin solution for 24 h, stored in 10% sucrose/formalin and sectioned via microtome. All brains processed for light microscopy using anti-tyrosine hydroxylase immunohistochemistry (AB152, Millipore-Sigma) and a NovaRed chromagen reaction (SK-4800, Vector Laboratories, Burlingame, CA). Sections were mounted, imaged using a light microscope (Axio Imager Z2, Zeiss, Thornwood, NY) and electrode placement was confirmed⁹⁴.

Moaddab M, & McDannald M.A. (2021). Retrorubral field is a hub for diverse threat and aversive outcome signals. Current biology : CB, 31(10), 2099-2110.e5.

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Pathology and Immunohistochemistry of Kangaroo and Mara

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The carcasses of a red kangaroo (Macropus rufus) and a Patagonian mara (Dolichotis patagonum) were submitted to the Department of Pathology, School of Veterinary Medicine, University of Chile for post mortem examination. A full necropsy was performed and in both animals, samples from the brain, trachea, lung, heart, liver, kidneys, spleen, stomach, small intestine, large intestine, adrenal gland and thyroid gland were collected. These samples were fixed in 10% buffered formalin, routinely processed for histopathology and stained with hematoxylin and eosin (H&E). Liver sections of the mara were also stained with Brown and Brenn Gram stain (CHURUKIAN & SCHENK, 1982) (link).
Immunohistochemistry for T. gondii detection, using samples of heart of both animals, was performed at the California Animal Health and Food Safety Laboratory, San Bernardino branch. Briefly, after antigen retrieval with pepsin (Sigma P7000; 0.4% HCl), rabbit anti-T. gondii polyclonal antibody (UCDavisRbt#58; 1:1500) was used, followed by anti-rabbit polymer (Dako K4003, Carpinteria, CA). The reaction was visualized with Novared for peroxidase following the manufacturer's instructions (VectorLab SK4800, Burlingame, CA). Slides were counterstained with Mayer hematoxylin, air dried and cover slipped. As a positive control, heart from a cat with a confirmed infection with T. gondii was used.

Díaz-Ayala N., Hidalgo-Hermoso E., Cabello-Araya C, & Carvallo-Chaigneau F. (2016). Infection with Toxoplasma gondii in a red kangaroo (Macropus rufus) and a Patagonian mara (Dolichotis patagonum) in captivity. Revista brasileira de parasitologia veterinaria = Brazilian journal of veterinary parasitology : Orgao Oficial do Colegio Brasileiro de Parasitologia Veterinaria, 25(4).

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