Appropriate serum

Manufactured by Agilent Technologies

Appropriate serum is a laboratory reagent used to maintain the appropriate balance of proteins, electrolytes, and other components in cell culture media. It provides a nutrient-rich environment to support the growth and maintenance of cells in in vitro experiments.

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Lab products found in correlation

Citrate solution, by Agilent Technologies (3 mentions) Bx41 microscope, by Olympus (2 mentions) Prolong gold mounting medium with dapi, by Thermo Fisher Scientific (1 mentions) Goat anti rabbit secondary antibody, by Jackson ImmunoResearch (1 mentions) D4w2z, by Cell Signaling Technology (1 mentions) Cellsense imaging software, by Olympus (1 mentions) Rat anti cd34, by BD (1 mentions) Rabbit anti phospho 4ebp1, by Cell Signaling Technology (1 mentions) Rat anti cd31, by BD (1 mentions)

3 protocols using appropriate serum

Immunohistochemical Staining for CD8+ T Cells

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For serial sections cut at 7-μm thickness, tissue samples were fixed in 2% paraformaldehyde overnight at 4°C, dehydrated, and embedded in paraffin. Paraffin slides were first rehydrated to proceed with antigen retrieval in citrate solution (Dako). Subsequently, the sections were blocked with the appropriate serum (Dako) and incubated overnight with the rabbit anti-CD8 antibody (D4W2Z; Cell Signaling Technology) at 1:200 dilution. Next, the secondary goat anti-rabbit antibody (Jackson Immunoresearch, West Grove, PA) was used at 1:300 dilution. Whenever sections were stained in fluorescence, ProLong Gold mounting medium with DAPI (Invitrogen, San Diego, CA) was used. Microscopic analysis was done with an Olympus BX41 microscope (Olympus, Tokyo, Japan) and CellSense imaging software V2.3 (Olympus Lifescience, Tokyo, Japan).

Serra M., Di Matteo M., Serneels J., Pal R., Cafarello S.T., Lanza M., Sanchez-Martin C., Evert M., Castegna A., Calvisi D.F., Mazzone M, & Columbano A. (2022). Deletion of Lactate Dehydrogenase-A Impairs Oncogene-Induced Mouse Hepatocellular Carcinoma Development. Cellular and Molecular Gastroenterology and Hepatology, 14(3), 609-624.

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Immunohistochemistry of Lung Tissue

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Immunohistochemistry on lung sections was performed as previously reported (Wenes et al., 2016 (link)). In brief, for serial sections of lungs from FASN cut at 7 μm thickness, tissue samples were fixed in 2% PFA overnight at 4°C, dehydrated and embedded in paraffin. Paraffin slides were first rehydrated to further proceed with antigen retrieval in citrate solution (DAKO). Sections were then fixed in 100% methanol. If necessary, 0.3% hydrogen peroxide was added to methanol, to block endogenous peroxidases. The sections were blocked with the appropriate serum (DAKO) and incubated overnight with the following antibodies: rabbit anti-FASN (Abcam ab 99359), rat anti-CD34 (BD Pharmingen), rabbit anti-phoshpo-p70S6K (T389) (Cell Signaling, 9205), rabbit anti-phospho-4EBP1 (Cell Signaling 2855. Appropriate biotin-labeled secondary antibodies (Jackson Immunoresearch) 1:300 were used, along with streptavidin-bound peroxidase. When necessary, TCA fluoresceine-tyramine or TSA Plus Cyanine 3 system amplification (Perkin Elmer, Life Sciences) were performed according to the manufacturer’s instructions. The sections were subsequently stained with Hoechst. ProLong Gold mounting medium without DAPI (Invitrogen) was used. Microscopic analysis was done with an Olympus BX41 microscope and CellSense imaging software.

Bruning U., Morales-Rodriguez F., Kalucka J., Goveia J., Taverna F., Queiroz K.C., Dubois C., Cantelmo A.R., Chen R., Loroch S., Timmerman E., Caixeta V., Bloch K., Conradi L.C., Treps L., Staes A., Gevaert K., Tee A., Dewerchin M., Semenkovich C.F., Impens F., Schilling B., Verdin E., Swinnen J.V., Meier J.L., Kulkarni R.A., Sickmann A., Ghesquière B., Schoonjans L., Li X., Mazzone M, & Carmeliet P. (2018). Impairment of Angiogenesis by Fatty Acid Synthase Inhibition Involves mTOR Malonylation. Cell metabolism, 28(6), 866-880.e15.

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Immunohistochemical Analysis of Tumor Vasculature

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For serial sections cut at 7 μm thickness, tumor samples were fixed in 2% PFA overnight at 4°C, dehydrated and embedded in paraffin. Paraffin slides were first rehydrated to further proceed with antigen retrieval in citrate solution (DAKO). The sections were blocked with the appropriate serum (DAKO) and incubated overnight with rat anti-CD31 (BD Pharmingen) 1:200. Appropiate secondary antibodies were added with Hoescht 33342 (Life thechnologies). ProLong Gold mounting medium was used. Microscopic analysis was done with an Olympus BX41 microscope and CellSense imaging software.

Martín-Pérez R., Yerbes R., Mora-Molina R., Cano-González A., Arribas J., Mazzone M., López-Rivas A, & Palacios C. (2017). Oncogenic p95HER2/611CTF primes human breast epithelial cells for metabolic stress-induced down-regulation of FLIP and activation of TRAIL-R/Caspase-8-dependent apoptosis. Oncotarget, 8(55), 93688-93703.

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