Tumors were fixed with Streck Cell Preservative (Streck, Omaha, NE, USA) for 48h and processed as paraffin-embedded tissue sections. Immunostaining was performed in accordance with the manufacturer's instructions (Vector, Versatile ABC, Burlingame, CA, USA). Briefly, tumor sections were deparaffinized in xylene and rehydrated in graded ethanol. Antigens were retrieved by maintaining tissue in 100 mM citrate buffer (Sigma-Aldrich, St Louis, MO, USA) (pH 6.0) at 95 °C for 30 min. Endogenous peroxidase was blocked by 3% H2O2 in 100% methanol for 10 min and nonspecific binding was reduced with 5% rabbit serum for 30 min at room temperature. For detection of luciferase, tumor sections were incubated with rabbit polyclonal antibody (targeting luciferase; dilution 1:500; Abcam, Cambridge, MA, USA) at 4 °C overnight. After incubation with a secondary anti-rabbit biotin-labeled antibody for 30 min and then with peroxidase-labeled streptavidin (Vector) for an additional 30 min, chromogen diaminobenzidine was added to tumor sections for 5 min for color development. The tumor sections were then dehydrated and mounted with glass coverslips. Three independent experiments were performed.
Cell preservative
Manufactured by Streck
Sourced in United States
Streck Cell Preservative is a reagent designed to preserve cellular components for analysis. It maintains the integrity of cells, allowing for accurate measurement and evaluation of cellular properties.
Automatically generated - may contain errors
Lab products found in correlation
Citrate buffer, by Merck Group (2 mentions)
Versatile abc, by Vector Laboratories (2 mentions)
Rabbit polyclonal antibody targeting luciferase, by Abcam (2 mentions)
Peroxidase labeled streptavidin, by Vector Laboratories (2 mentions)
Xt 2000iv, by Sysmex (1 mentions)
Reticulocyte stain, by Merck Group (1 mentions)
Edta vacutainer, by BD (1 mentions)
4 protocols using cell preservative
1
Immunohistochemical Analysis of Luciferase Expression
2
Immunohistochemical Detection of Luciferase
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Tumors were fixed with Streck Cell Preservative (Streck, Omaha, NE, USA) for 48 h and processed as paraffin-embedded tissue sections. Immunostaining was performed in accordance with the manufacturer's instructions (Vector, Versatile ABC, Burlingame, CA, USA). Briefly, tumor sections were deparaffinized in xylene and rehydrated in graded ethanol. Antigens were retrieved by maintaining tissue in 100 mM citrate buffer (Sigma-Aldrich, St Louis, MO, USA) (pH 6.0) at 95 °C for 30 min. Endogenous peroxidase was blocked by 3% H2O2 in 100% methanol for 10 min and nonspecific binding was reduced with 5% rabbit serum for 30 min at room temperature. For detection of luciferase, tumor sections were incubated with rabbit polyclonal antibody (targeting luciferase; dilution 1:500; Abcam, Cambridge, MA, USA) at 4 °C overnight. After incubation with a secondary anti-rabbit biotin-labeled antibody for 30 min and then with peroxidase-labeled streptavidin (Vector) for an additional 30 min, chromogen diaminobenzidine was added to tumor sections for 5 min for color development. The tumor sections were then dehydrated and mounted with glass coverslips. Three independent experiments were performed.
3
Comprehensive Hematological Analysis of Neonates
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Packed cell volume (PCV; L/L) and total plasma protein (TPP; g/L) concentration were manually determined on the day of sampling, while automated hematological analysis (Sysmex XT-2000iV, Sysmex, Kobe, Japan) of preserved blood specimens (Streck Cell Preservative; Streck, Omaha, USA) was performed 2–8 days later (21 (link)). Automated analysis provided indicators of red blood cell number additional to PCV (red blood cell [RBC; ×10^12/L] count and hemoglobin [Hb; g/L] concentration), as well as platelet (PLT; ×10^9/L) and total white blood cell (WBC; ×10^9/L) counts. Duplicate Wrights-Giemsa-stained blood smears prepared the day of sampling were later reviewed for manual differential WBC and nucleated RBC (nRBC) counts. Reticulocyte percentage was determined from new methylene blue-stained blood smear counts (Reticulocyte stain, Sigma-Aldrich, Germany) using a Miller Square (9:1 ratio) eyepiece reticule to count the equivalent of >1,000 RBC. In addition to nRBC, the absolute reticulocyte count for each pup was subsequently calculated (reticulocyte percentage x RBC count) as a measure of the intensity of RBC regeneration.
4
Blood Preservation and Storage
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The venous blood was drawn into lithium heparin (LH) or EDTA Vacutainers [Becton Dickinson (BD); USA]. Within 2 h after blood collection, a 1 mL aliquot of the human blood specimen was added to 1 mL of the Streck Cell Preservative (Catalogue # 213350; Streck, USA). The sample was mixed by pipetting. A 1 mL aliquot of both the Streck Cell Preserved-specimen and original whole blood specimens were stored at 4 C refrigerator and at RT.
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