Immunocytochemical procedures were performed as described previously50 (link). Briefly, cells were collected by pipetting in PBS, plated on coverslips coated with poly-L-lysine (Sigma-Aldrich) and fixed. Cells were concurrently incubated with primary antibodies (rabbit anti-CD133, Abcam; goat anti-rat CD34, R&D Systems) at 4 °C overnight. The nuclei of cells were counterstained with DAPI (Sigma).
Rabbit anti cd133
Manufactured by Abcam
Sourced in United States, United Kingdom
Rabbit anti-CD133 is a primary antibody that detects the CD133 protein. CD133 is a cell surface glycoprotein that is commonly used as a marker for stem and progenitor cells.
Automatically generated - may contain errors
Lab products found in correlation
Poly l lysine (pll), by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Rabbit anti ebf1 polyclonal antibody, by Abcam (1 mentions)
Envision secondary antibody, by Agilent Technologies (1 mentions)
Dab substrate kit, by Vector Laboratories (1 mentions)
Paraformaldehyde (pfa), by BD (1 mentions)
Ab16287, by Abcam (1 mentions)
A1 confocal microscope, by Nikon (1 mentions)
Rat anti cd45, by Abcam (1 mentions)
Sudan black b, by Merck Group (1 mentions)
Goat anti rat igg, by Abcam (1 mentions)
Goat anti rabbit igg, by Abcam (1 mentions)
Goat anti mouse igg, by Abcam (1 mentions)
Alexa 488 conjugated goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Apc conjugated mouse anti cd44, by BD (1 mentions)
Pe conjugated mouse anti cd133, by Miltenyi Biotec (1 mentions)
Mouse anti o glcnac rl2, by Thermo Fisher Scientific (1 mentions)
Rabbit anti β tubulin, by Cell Signaling Technology (1 mentions)
Rabbit anti cd44, by Abcam (1 mentions)
Alexa fluor 488 donkey anti goat, by Thermo Fisher Scientific (1 mentions)
9 protocols using rabbit anti cd133
1
Immunocytochemical Characterization of CD133 and CD34
2
Immunohistochemical Analysis of EBF1 and CD133
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MMNK1 (60,000 cells/well), ox-MMNK1-L (60,000 cells/well), CCA cell lines (30,000 cells/well) were placed on 48-well plates for overnight. The cells were fixed with 4% paraformaldehyde-containing PBS for 30 min at room temperature. After washing, 0.2% (v/v) Triton-X100 solution was added. The cells were washed with PBS and incubated for 30 min with PBS containing 0.3% (v/v) hydrogen peroxide for endogenous hydrogen peroxide activity blocking and non-specific binding was blocked by 3% (w/v) BSA in PBS for 30 min. Cells were incubated with 10 µg/ml of rabbit anti-EBF1 polyclonal antibody (Abcam, MA, USA) or 9 µg/ml of rabbit anti-CD133 (Abcam, MA, USA) at room temperature for overnight followed by peroxidase-conjugated Envision™ secondary antibody (DAKO, Glostrup, Denmark). The color was developed with DAB substrate kit (Vector Laboratories, Inc., CA, USA) and washed with distilled water. The stained cells were dehydrated with stepwise (5 min/step) increasing concentrations of ethanol (70%→80%→90%→100%) and air dried overnight. The stained cells were examined under an inverted microscope.
3
Immunohistochemical Characterization of Cardiac Cells
4
Investigating Cancer Stem Cell Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used in the experiments: allophycocyanin (APC)-conjugated mouse anti-CD44 from BD Biosciences, phycoerythrin (PE)-conjugated mouse anti-CD133 from Miltenyi Biotec, rabbit anti-CD44, rabbit anti-CD133, and mouse anti-OGT from Abcam, Alexa 647-conjugated rabbit anti-mouse from Invitrogen; rabbit anti-β-tubulin from Cell Signaling Technology (Danvers, MA, USA); Alexa 488-conjugated goat anti-rabbit from Molecular Probes, Inc., (Eugene, OR, USA), mouse anti-O-GlcNAc (RL2) from Thermo Fisher Scientific; mouse anti-GAPDH from Santa Cruz Biotechnoloigy Inc (Sta. Cruz, CA, USA).
5
Immunofluorescence staining of cancer stem cell markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunofluorescence staining on tissue sections, CCSCs, mCCSCs and cCCSCs cells antibodies/antisera used were: mouse anti-Human Nuclei (1:100, Millipore-Merk KGaA, Darmstadt, Germany), mouse anti-BMI1 (1:100, Merk Millipore), rabbit anti-EpCAM (1:50, Cell Signaling, Beverly, MA, USA), rabbit anti-ALDHA1 (1:50, Cell Signaling), rabbit anti-bCatenin (1:50, Cell Signaling), rabbit anti-CD133 (1:100, AbCam, Cambridge, UK), rabbit anti-OLFM4 (1:200, AbCam), rabbit anti-CDX2 (1:100, AbCam), rabbit anti-CK20 (1:100, Abcam), rabbit anti-Lgr5 (1:50, Merk SigmaAldrich), rabbit anti-Laminin (1:400, Merk SigmaAldrich), mouse anti-Vimentin (1:100, Agilent, Santa Clara, CA, USA), mouse anti-HLA-ABC (1:100, Agilent), goat anti-EphA2 (1:50, R&D System, Minneapolis, MN, USA), mouse anti-CXCR4 (1:100, R&D System), rabbit anti-KI67 (1:200, Leica Microsystem, Wetzlar, Germany), mouse anti-CD44 (1:50, BD Biosciences, San Jose, CA, USA), rabbit anti-Wnt5a (1:50, LS Biosciences, Seattle WA, USA), mouse anti-MUC2 (1:50, Novusbio, Littleton, CO, USA). Goat anti mouse AlexaFluor488/546 (1:1000, Thermo Fisher Scientific, Waltham, MA, USA), goat anti rabbit AlexaFluor488 (1:1000, Thermo Fisher Scientific) and donkey anti goat AlexaFluor488 (1:1000, Thermo Fisher Scientific) secondary antibodies were used to visualize the primary antibody staining.
6
Characterization of Schwann Cell Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Monolayer-cultured rBM-NCPs were fixed with 4% (wt/vol) paraformaldehyde, blocked, and then incubated with primary antibodies, including rabbit-anti-CD133 (1:200, Abcam), rabbit-anti-p75 (1:1000, Cell Signaling), rabbit-anti-nestin (1:50, Sigma), mouse-anti-vimentin (1:25, Sigma), mouse-anti-CD29 (1:100, Sigma), and rabbit-anti-Ki67 (1:200, Sigma), overnight at 4 °C, followed by reaction with FITC-anti-rabbit-IgM (Sigma), Cy3-anti-rabbit-IgM (Santa Cruz), TRITC-anti-mouse-IgM (Santa Cruz) or FITC-anti-mouse-IgM (Sigma), and Hoechst 33342 (Sigma) counterstaining. The cell samples were observed under a confocal laser scanning microscope (TCS SP2, Leica Microsystems, Germany).
Induced Schwann cells were subjected to immunofluorescent staining with mouse anti-S100β (1:250, Sigma), rabbit-anti-glial fibrillary acidic protein (anti-GFAP, 1:200, DakoCytomation), and rabbit-anti-p75 (1:1000, Cell Signaling Technology) respectively, followed by the same reaction with the second antibody and Hoechst 33342 counterstaining.
Induced Schwann cells were subjected to immunofluorescent staining with mouse anti-S100β (1:250, Sigma), rabbit-anti-glial fibrillary acidic protein (anti-GFAP, 1:200, DakoCytomation), and rabbit-anti-p75 (1:1000, Cell Signaling Technology) respectively, followed by the same reaction with the second antibody and Hoechst 33342 counterstaining.
7
Histological and Immunofluorescence Analysis of Kidney Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples were prepared for histological and immunofluorescence analyses by following standard protocols for paraffin embedding. For histological analysis, mounted kidney sections were stained by hematoxylin and eosin.
For immunofluorescence staining, the sections were blocked with 5% normal bovine serum in PBS for 30 min. Sections were then incubated with primary antibodies overnight at 4°C. The primary antibodies used were as follows: mouse anti-VEGFR2 (1:200; Abcam, UK), rabbitanti-CD133 (1:200; Abcam), goat anti-fibronectin (1:200; Abcam) and rabbit anti-CD31 (1:200; Abcam). After being washed in PBS, sections were then incubated in 488- or 594-conjugated species-specific secondary antibody (1:100; Chemicon, USA) for 2 hours. Slides were observed with an Olympus fluorescent microscope and images were captured using Olympus soft image viewer. The intensity of immunoreactivity was quantified using the Image-Pro Plus 6.0 software (Media Cybernetics, USA).
For immunofluorescence staining, the sections were blocked with 5% normal bovine serum in PBS for 30 min. Sections were then incubated with primary antibodies overnight at 4°C. The primary antibodies used were as follows: mouse anti-VEGFR2 (1:200; Abcam, UK), rabbitanti-CD133 (1:200; Abcam), goat anti-fibronectin (1:200; Abcam) and rabbit anti-CD31 (1:200; Abcam). After being washed in PBS, sections were then incubated in 488- or 594-conjugated species-specific secondary antibody (1:100; Chemicon, USA) for 2 hours. Slides were observed with an Olympus fluorescent microscope and images were captured using Olympus soft image viewer. The intensity of immunoreactivity was quantified using the Image-Pro Plus 6.0 software (Media Cybernetics, USA).
8
Immunofluorescence Staining of Stem Cell Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cultured DP cells were fixed using 4% (w/v) paraformaldehyde fixation for 10 min. The cells were permeabilized for 5 min using PBS with 0.3% Triton X-100 (T8787, Sigma). Blocking was performed for 30 min using 3% (w/v) bovine serum albumin in PBS. The primary antibodies including rabbit anti-CD133 (1:500; Abcam, Cambridge, MA, USA), mouse anti-SOX2 (1:200; SantaCruz, Santa Cruz, CA, USA) and rabbit anti-Versican (1:500; Bioss) left overnight at 4 °C. The next day, the primary antibodies were washed off using three consecutive PBS washes. The secondary antibody, Alexafluor 594 goat anti-rabbit (1:1000; Thermo, Agawam, MA, USA)/Alexafluor 488 rabbit anti-mouse (1:1000; Thermo) was applied for 1 h at room temperature. The nuclei of cells were counterstained with DAPI (C1005, Beyotime) for 5 min at room temperature after removing the secondary antibody using three consecutive PBS washes. The coverslips were mounted using anti-fade reagent and the cells were visualized on a Zeiss Exciter fluorescent microscope. The specific information on all the antibodies used is listed in Supplementary Table S1 .
9
Immunostaining of Neural Stem Cell Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were xed 4% paraformaldehyde in 0.2M phosphate-buffered saline (PBS) for 20 min, were permeabilized in 0.3% Triton X-100 in PBS for 5 min and blocked with 10% normal goat serum for 10 min at room temperature, respectively. Then, cells were incubated with the appropriate antibody overnight at 4°C. The primary antibodies were rabbit anti-nestin (1:250; Abcam), rabbit anti-CD133 (1:500; Abcam), rabbit anti-GFAP (1:300, Abcam), rabbit anti-MAP2 (1:300, Abcam), rabbit anti-OLIG2 (1:300, Abcam). Following washing with PBS, the cells were incubated with secondary FITC-conjugated goat anti rabbit antibody (1:1000; Abcam) and FITC-conjugated goat anti mouse antibody (1:1000; Abcam) for 2 h at room temperature. Nuclei were stained with DAPI (blue). Finally, immunopositive cells were observed using inverted uorescence microscopy.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!