A total of 50,000 unsorted or FACS sorted cells from the different cell lines were cytospun and stained. Paraffin embedded xenograft tumor blocks were cut into 6 μm sections and immune-stained as described 25 (link). The primary antibodies used were CD44v, panCD44, CD34 (Abcam), e-cadherin (BD), EGFR (Santa Cruz), pEGFR (BD), xCT 20 (link) and PD-L1 (Novus Bio). The appropriate Alexa Fluor-coupled secondary antibodies were used in double staining sections. Nuclei were stained with DAPI (Vector labs - Burlingame, CA). Slides were examined by fluorescent microscopy using a Zeiss AxioImager microscope (Carl Zeiss, Jena, Germany).
Pancd44
Manufactured by Abcam
PanCD44 is an antibody that recognizes the CD44 antigen, a cell surface glycoprotein involved in cell-cell interactions, cell adhesion, and migration. This antibody can be used to detect the expression of CD44 in various cell types and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Pd l1, by Novus Biologicals (1 mentions)
Axioimager microscope, by Zeiss (1 mentions)
Pegfr, by BD (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions)
E cadherin, by BD (1 mentions)
Bicinchoninic acid protein assay kit, by Thermo Fisher Scientific (1 mentions)
Glut1, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
2 protocols using pancd44
1
Immunofluorescence Analysis of Tumor Xenografts
2
Protein Expression Analysis in Cells
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RIPA buffer (Beyotime) was used for protein extraction. After the total protein concentration was determined by a bicinchoninic acid protein assay kit (Thermo Fisher Scientific, Inc), 20 μg protein samples were separated by 8% SDS polyacrylamide gels and transferred onto PVDF membranes (Millipore). The membranes were blocked with 5% skimmed milk in 0.1% TBS-Tween-20 at room temperature for 1 h and incubated with following primary antibodies overnight at 4 °C: CD44v10 (1:1000, Sigma-Aldrich, AB2082), pan-CD44 (1:1000, Abcam, ab189524), GLUT1 [1:1000, Cell Signaling Technology, (CST), 73,015], Oct4 (1:1000, CST, 2750), c-Myc (1:1000, CST, 5605), Klf4 (1:1000, CST, 4038), ALDH1 (1:1000, CST, 12,035), Nanog (1:1000, CST, 4903), p-ERK (1:2000, CST, 4370), ERK (1:1000, CST, 4695), p-AKT (1:1000, CST, 4060), AKT (1:1000, CST, 4691), and β-actin (1:1000, CST, 3700). On the following day, horseradish peroxidase-conjugated secondary antibodies (1:5000, Lianke Biotech Co., Ltd) were added. Then, the protein bands were subsequently captured using the enhanced plus chemiluminescence assay (Pierce) and were measured on an ImageQuant LAS 4000 mini.
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