TDCs were seeded at a cell density of 5 × 104 cells/well and incubated at 37 °C at 5% CO2. After 24 h incubation, cells were fixed at 37 °C for 10 min with 4% formaldehyde (pH 7.4). They were then washed twice with PBS and permeabilized at room temperature for 10 min. Next, cells were blocked in blocking solution (2% bovine serum albumin in PBS) for 1 h at room temperature, incubated overnight with primary antibodies diluted in blocking solution at 4 °C, washed three times for 5 min with PBS, incubated with secondary antibody at RT for 2 h, and washed four times for 5 min with PBS. The primary antibodies used for immunostaining and their dilutions were as follows: rabbit anti-tenomodulin (1:50, Abcam, Cambridge, UK), mouse anti-CD44 (1:50, BD Biosciences), mouse anti-90(1:50, BD Biosciences). The secondary antibodies used were Alexa Fluor 488-conjugated goat anti-mouse IgG (1:100, Thermo, Waltham, US) and Alexa Fluor 555-conjugated goat anti-rabbit IgG (1:100, Thermo).
Mouse anti cd44
Manufactured by BD
Sourced in United Kingdom, United States, Germany
Mouse anti-CD44 is a laboratory reagent used to detect and quantify the presence of the CD44 protein in cellular samples. CD44 is a cell surface glycoprotein involved in cell-cell interactions, cell adhesion, and migration. This product can be used in various immunoassay techniques to study the expression and distribution of CD44 in different cell types and biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 555 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Rabbit anti tenomodulin, by Abcam (1 mentions)
Alexa fluor 488 conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Axioplan 2 microscope, by Leica (1 mentions)
Dm ire2 confocal microscope, by Leica (1 mentions)
Mouse anti cxcr4, by R&D Systems (1 mentions)
Rabbit anti cdx2, by Abcam (1 mentions)
Goat anti epha2, by R&D Systems (1 mentions)
Rabbit anti cd133, by Abcam (1 mentions)
Alexa fluor 488 donkey anti goat, by Thermo Fisher Scientific (1 mentions)
Rabbit anti β catenin, by Cell Signaling Technology (1 mentions)
Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
3 protocols using mouse anti cd44
1
Immunofluorescence Staining of Tenomodulin, CD44, and CD90
2
Immunohistochemical Analysis of Glioblastoma
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Example 2
Immunohistochemistry
Tissue samples from hGBM, normal human and mouse brain were post-fixed in 4% paraformaldehyde (PFA) for 24 h and placed in a sucrose solution at decreasing concentrations beginning at 30%. Hematoxylin and Eosin (H&E) staining and immunohistochemistry were performed on OCT-embedded, 10 μm-thick cryostat sections (Galli et al., 2004; Vescovi et al., 1999). Tissue sections were stained overnight at 4° C. with the following primary antibodies diluted in 10% normal goat serum (NGS; Gibco, Rockville, Md., USA): mouse anti-EphA2 cloneD7 (1:200; Sigma; St. Louis, Mo., USA), mouse anti-SSEA1 and mouse anti-CD44 (1:100; BD Biosciences, Franklin Lakes, N.J., USA). Goat anti-mouse AlexaFluor488/546 (1:2000; Invitrogen Corp, Carlsbad, Calif., USA) was then employed. Cell nuclei were counterstained by TO-PRO-3 (Molecular Probes, Invitrogen). Negative controls were obtained by omitting primary antibody. Samples were photographed with Zeiss Axioplan2 Microscope and Leica DMIRE2 Confocal Microscope.
3
Immunofluorescence staining of cancer stem cell markers
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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.
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