Cell lysates were prepared by adding two times Laemmli sample buffer to sorted CD11b+Gr-1+Sca-1− and CD11b+Gr-1+Sca-1+ myeloid cells. The protein samples were resolved on 10 or 12% SDS–polyacrylamide gel electrophoresis gels and transferred to polyvinylidene difluoride membranes. Membranes were blocked in 4% bovine serum albumin in TBS-T for 1 hour at room temperature. Primary antibodies used were anti–Sca-1 (1:2000) from Abcam (Cambridge, MA, USA) and anti–β-actin (1:2500) from Cell Signaling Technology (Beverly, MA, USA). Secondary horseradish peroxidase–conjugated antibodies used were anti-mouse IgG and anti-rabbit IgG (all 1:5000) from Enzo Lifesciences (Farmingdale, NY, USA).
Anti sca 1
Manufactured by Abcam
Sourced in United States
Anti-Sca-1 is a laboratory reagent used in flow cytometry and cell sorting applications. It is an antibody that specifically binds to the Sca-1 (Stem Cell Antigen-1) protein expressed on the surface of certain types of stem cells and progenitor cells. The primary function of Anti-Sca-1 is to enable the identification and isolation of these cell populations for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Cell Signaling Technology (1 mentions)
Cardiac troponin t, by Abcam (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Alpha sarcomeric actin, by Merck Group (1 mentions)
Zenon technology, by Thermo Fisher Scientific (1 mentions)
Aspirin, by Merck Group (1 mentions)
Wild type sprague dawley rats, by Charles River Laboratories (1 mentions)
Prasugrel, by Merck Group (1 mentions)
Anti ki67, by Santa Cruz Biotechnology (1 mentions)
Mouse c reactive protein crp elisa kit, by R&D Systems (1 mentions)
C kit, by Santa Cruz Biotechnology (1 mentions)
Anti nkx2, by Abcam (1 mentions)
Anti sca 1, by Santa Cruz Biotechnology (1 mentions)
Anti ki67, by Abcam (1 mentions)
Anti cd105, by Abcam (1 mentions)
Anti oct4, by Santa Cruz Biotechnology (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Anti cd31, by Abcam (1 mentions)
Ticagrelor, by AstraZeneca (1 mentions)
3 protocols using anti sca 1
1
Western Blot Analysis of Myeloid Cell Subsets
2
Multicolor Immunostaining of Cardiac Stem Cells
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After rinsing with PBS and fixation for 20 min in 2.5% paraformaldehyde at room temperature, the cells were permeabilized with 0.25% Triton-X100 for 10 min. For immunostaining we used primary mouse anti-Isl-1 antibodies (Abcam) and mouse antibodies to sarcomeric alpha-actinin (Abcam), alpha-sarcomeric actin (Sigma-Aldrich) and cardiac troponin T (Abcam) pre-conjugated with Alexa 532, 546, 568, 594, or 647 according to Zenon technology (Invitrogen). Phycoerythrin-conjugated anti-Ki-67 antibodies were used for determination of proliferative capacity of CSCs. Commercially available FITC-conjugated anti-c-kit (Abcam) and anti-Sca-1 (Abcam) antibodies were also used at 1:100 dilution. Hoechst 33342 (10 μg/mL, Molecular Probes, USA) staining at 1:1000 dilution was used for detection of cell nuclei.
Confocal microscope Leica TCS SP5 provides an opportunity to simultaneously analyze several fluorescent tags in wide spectrum (from ultraviolet to infrared), thus it is possible to use several fluorochromes and visualize expression of CSCs markers of several types at a single sample. Intensity of the corresponding fluorochrome serves to establish a type of CSC which generated a colony or CICS.
Confocal microscope Leica TCS SP5 provides an opportunity to simultaneously analyze several fluorescent tags in wide spectrum (from ultraviolet to infrared), thus it is possible to use several fluorochromes and visualize expression of CSCs markers of several types at a single sample. Intensity of the corresponding fluorochrome serves to establish a type of CSC which generated a colony or CICS.
3
Rat Cardiac Regeneration Evaluation
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Wild-type Sprague-Dawley rats were purchased from Charles River Laboratories (Wilmington, MA). Ticagrelor was provided by AstraZeneca. Aspirin and prasugrel were purchased from Sigma.
Mouse C-Reactive Protein/CRP ELISA Kit was purchased from R&D systems. Myocardial PGE 2, 6-keto-PGF 1α and 15-epi-lipoxin A 4 ELISA kits were purchased from Cayman Chemical. TRIzol reagent was purchased from Invitrogen, Carlsbad, CA. Anti-Ki-67, c-Kit, anti-Oct4 and anti-Sca-1 antibodies for the immunoblotting were purchased from Santa Cruz Biotechnology and anti-β-actin from Sigma-Aldrich. Anti-CD31, Anti-Ki-67, anti-CD105, anti-Sca-1 and anti-Nkx2.5 antibodies for the immunohistochemistry were purchased from Abcam. Anti-CD 31 http://www.abcam.com/cd31-antibody-tld-3a12-ab64543. htmland anti-c-Kit antibodies for the immunofluorescence staining were purchased from Abcam and the fluorescence-labeled secondary antibodies (AlexaFluor 488 for CD31 and AlexaFluor 546 for c-Kit) were purchased from Invitrogen.
Mouse C-Reactive Protein/CRP ELISA Kit was purchased from R&D systems. Myocardial PGE 2, 6-keto-PGF 1α and 15-epi-lipoxin A 4 ELISA kits were purchased from Cayman Chemical. TRIzol reagent was purchased from Invitrogen, Carlsbad, CA. Anti-Ki-67, c-Kit, anti-Oct4 and anti-Sca-1 antibodies for the immunoblotting were purchased from Santa Cruz Biotechnology and anti-β-actin from Sigma-Aldrich. Anti-CD31, Anti-Ki-67, anti-CD105, anti-Sca-1 and anti-Nkx2.5 antibodies for the immunohistochemistry were purchased from Abcam. Anti-CD 31 http://www.abcam.com/cd31-antibody-tld-3a12-ab64543. htmland anti-c-Kit antibodies for the immunofluorescence staining were purchased from Abcam and the fluorescence-labeled secondary antibodies (AlexaFluor 488 for CD31 and AlexaFluor 546 for c-Kit) were purchased from Invitrogen.
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