When animals were moribund, brains were collected and fixed in 10% formalin, sectioned in 4 micron sections and immunostained with antibodies to cleaved caspase 3 (CP229C, BioCare Medical, Concorde, CA, 1/500), Ki67 (TA-125-MH, ThermoShandon, Fremont, CA, 1:500), and Abcg2 (BXP53, Enzo Life Sciences, Farmingdale, NY, 1:10,000).
Cp229c
Manufactured by Biocare Medical
Sourced in United States
The CP229C is a centrifuge product manufactured by Biocare Medical. It is designed to separate components of liquid samples through centrifugal force. The device can accommodate various sample volumes and speeds to facilitate sample preparation for further analysis.
Automatically generated - may contain errors
Lab products found in correlation
Bxp 53, by Enzo Life Sciences (1 mentions)
Ventana discovery ultra, by Roche (1 mentions)
Eclipse ni microscope, by Nikon (1 mentions)
Superfrost plus, by Thermo Fisher Scientific (1 mentions)
Discovery chromomap dab kit, by Roche (1 mentions)
Omnimap anti rabbit hrp, by Roche (1 mentions)
Sc 1348, by Santa Cruz Biotechnology (1 mentions)
Ab76003, by Abcam (1 mentions)
Sc 7764, by Santa Cruz Biotechnology (1 mentions)
4 protocols using cp229c
1
Quantifying Apoptosis and Proliferation
2
Histopathological Analysis of Hepatic Tissues
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Hepatic tissues were preserved in 10% formalin. They were then routinely processed until being embedded in paraffin blocks. Sections were cut to a 5 μm thickness andprocessed until being stained with hematoxylin and eosin according to Bancroft and Layton [32 ]. For immunohistochemical staining of the hepatic sections, the primary antibody against caspase-3 (polyclonal rabbit anti-cleaved caspase-3 at dilution 1:100, BioCare Medical, Cat: CP229C, Concord, CA, USA) was utilized. Immunostaining intensity was quantified using Image J software (National Institutes of Health, Bethesda, MD, USA) [33 (link)].
3
Apoptosis Quantification in Xenografts
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Xenografts were fixed in 10% neutral buffered formalin, embedded in paraffin, sectioned at 4 μM, stained with hematoxylin and eosin and reviewed by light microscopy using an upright Nikon Eclipse Ni microscope (Nikon Instruments, Inc.). Immunohistochemistry was performed on 4 μm thick tissue sections mounted on positively charged glass slides (Superfrost Plus; Thermo Fisher Scientific, Waltham, MA), and dried at 60°C for 20 min. The procedures for immunohistochemistry were performed using a Ventana DISCOVERY ULTRA autostainer (Roche). Analysis of cleaved caspase 3 IHC . Immunohistochemistry for cleaved caspase 3 (BioCare medical, CP229C, 1:500) was performed using a Ventana DISCOVERY ULTRA automated stainer (Ventana Medical Systems, Inc., Tucson, AZ). Heat-induced epitope retrieval (HIER) was carried out for 32 min at 37°C using Cell conditioning media 1 (CC1, 950-124, Ventana Medical Systems, Inc, Tucson, AZ) followed by incubation with the primary antibody for 60 min and then visualization with OmniMap anti-rabbit HRP (760–4311, Ventana Medical Systems, Inc, Tucson, AZ) and the DISCOVERY ChromoMap DAB kit (760-159, Ventana Medical Systems, Inc, Tucson, AZ). Positive cells in different views under microscope were counted as individuals for unpaired Student’s t test.
4
Immunohistochemical Analysis of Distal Femur
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Sections of the distal femur were adhered to silanized slides and submitted to immunohistochemistry for anti-Caspase-3 (Cleaved) (CP-229C, 1:30, Biocare Medical, Concord, Ca), anti-type II collagen (sc-7764, 1:250, Santa Cruz Biotechnology), anti-TNF-a (sc-1348, 1:200, Santa Cruz Biotechnology) and anti-MMP-9 (ab76003, 1:100, Abcam) according to our previous protocol 35 . The percentage of positive chondrocytes for caspase-3, tumor necrosis factor-a (TNF-a) and MMP-9 were calculated using the number of immune-positive cells divided by the total cell number from a predetermined rectangular area of 40.000 mm 2 in WB and NWB areas.
To analyze type II collagen expression in superficial zone (SZ), intermediate zone (IZ) and deep zone (DZ) from WB and NWB areas, two sections from three animals/group (200Â magnification) were used, and 60 different points were analyzed in each area to calculate the average of the densitometric grey levels using an image analysis system (AxioVision Rel. 4.9., Carl Zeiss, Germany) 35 .
To analyze type II collagen expression in superficial zone (SZ), intermediate zone (IZ) and deep zone (DZ) from WB and NWB areas, two sections from three animals/group (200Â magnification) were used, and 60 different points were analyzed in each area to calculate the average of the densitometric grey levels using an image analysis system (AxioVision Rel. 4.9., Carl Zeiss, Germany) 35 .
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