alcam polyclonal antibody  (Proteintech)

Journal: Development (Cambridge, England)

Article Title: Core Hippo pathway components act as a brake on Yap and Taz in the development and maintenance of the biliary network

doi: 10.1242/dev.184242

Figure Lengend Snippet: Hepatocyte apical domains appear abnormal and expanded in sav1−/− larvae. (A-B‴) Examples of wild-type (A) and sav1−/− (B-B‴) 8 dpf larvae whole-mount immunostaining for Alcam. n=5 wild type, n=6 sav1−/−. Scale bar: 100 μm. (C,D) Single plane images of intrahepatic biliary ducts stained for annexin A4 in wild-type and sav1−/− 8 dpf larvae respectively. Red arrow in C indicates one annexin A4+ biliary epithelial cell. (C′,D′) Single plane images of aPKC staining in wild-type and sav1−/− 8 dpf larvae, respectively. Yellow arrow in C′ indicates one canaliculus. (C″,D″) Merged images with green representing aPKC and magenta representing annexin A4. Scale bar: 25 μm. (E-F″) Examples of wild-type and sav1−/− gallbladder stained for aPKC and annexin A4, following the same labeling conventions as in C-D″. (G,G′) Z-projections of whole-mount staining for annexin A4 and aPKC in wild-type larvae at 8 dpf. Scale bar: 25 μm. (H,H′,I,I′,J,J′) Z-projections of whole-mount staining for annexin A4 and aPKC in sav1−/− larvae at 8 dpf. (G″,H″,I″,J″) Merged images with green representing aPKC and magenta representing annexin A4 (n=7 wild type; n=5 sav1−/−). Scale bar: 100 μm. (K-K″) Z-projections of whole-mount staining for annexin A4 and Cdh2 in wild-type larvae at 8 dpf. (L,L′,M,M′,N,N′) Z-projections of whole-mount staining for annexin A4 and Cdh2 in sav1−/− larvae at 8 dpf. Yellow arrowheads in N′ indicate examples of high intensity cytoplasmic punctate Cdh2 staining, suggesting cadherin junction collapse. (L″,M″,N″) Merged images of Cdh2 (green) and annexin A4 (magenta) (n=7 wild type; n=6 sav1−/−). Scale bar: 100 μm.

Article Snippet: The following primary antibodies and concentrations were used: 1:200 dilution rabbit monoclonal Yap (ab81183, Abcam), 1:200 dilution rabbit monoclonal Yap and Taz (D24E4, Cell Signaling, 8418), 1:500 dilution rabbit monoclonal GFP ( {"type":"entrez-nucleotide","attrs":{"text":"G10362","term_id":"942211","term_text":"G10362"}} G10362 , ABfinity), 1:100 dilution mouse polyclonal Alcam (zn-8, DSHB), 1:200 dilution mouse monoclonal AnnexinA4/2F11 [ab71286, Zebrafish Gut Secretory Cell Epitope (FIS2F11/2)], 1:500 dilution rabbit polyclonal aPKC λ and ξ (C-20 and sc-216, Santa Cruz Biotech), 1:200 dilution rabbit polyclonal Cdh2 (GTX125962, GeneTex) and 1:500 dilution rabbit polyclonal Prox1 (AB5475, EMD Millipore).

Techniques: Immunostaining, Staining, Labeling

Journal: Journal of Translational Medicine

Article Title: Comparison of functional characterization of cancer stem cells in different tumor tissues of pseudomyxoma peritonei

doi: 10.1186/s12967-024-05730-6

Figure Lengend Snippet: CSCs sorting process. A . Flow cytometry was used to detect CD133 + , CD166 + , and CD133 + /CD166 + CSCs populations within mixed cells, with the R2 gate serving as the target cell population. B . Sorting of CSCs from three tissue classes using the cartridge of the MACSQuant Tyto system, with the cartridge and microchip on the left and cell sorting through the microchip on the right. C . Real-time visualization of CSCs obtained by FlowSight imaging flow cytometer. Bright field and cell fluorescence image are presented, with Ch01: light field, Ch03: CD133-PE, and Ch11: CD166-BV421; scale bar = 20 μm. Flow cytometric analysis was performed on the sorted negative cells

Article Snippet: To identify the CSC population by immunofluorescence staining, a polyclonal rabbit anti-CD133 antibody (1:200, Cat. #SAB5701045, Sigma-Aldrich, St. Louis, Missouri, USA) and polyclonal mouse anti-CD166 antibody (1:50, Cat. #AF1172, R&D Systems, Minneapolis, Minnesota, USA), were employed.

Techniques: Flow Cytometry, MicroChIP Assay, FACS, Imaging, Fluorescence

Journal: Journal of Translational Medicine

Article Title: Comparison of functional characterization of cancer stem cells in different tumor tissues of pseudomyxoma peritonei

doi: 10.1186/s12967-024-05730-6

Figure Lengend Snippet: Cell culture and marker identification. A . Observation of cell growth using a light microscope revealed the morphology, size, and adherence of cells in the logarithmic growth phase, with a scale bar indicating 100 μm. B . Immunofluorescence staining was performed to detect markers. Nuclei are stained blue, CD133 + appears as green fluorescence, CD166 + appears as red fluorescence, and CD133 + /CD166 + appears as yellow fluorescence. Scale bar indicates 200 μm. C . Marker detection was also conducted using flow cytometry, where each colored point represents a cell. The target cell population is enclosed by a black box for clarity, with brighter colors indicating a higher cell density

Article Snippet: To identify the CSC population by immunofluorescence staining, a polyclonal rabbit anti-CD133 antibody (1:200, Cat. #SAB5701045, Sigma-Aldrich, St. Louis, Missouri, USA) and polyclonal mouse anti-CD166 antibody (1:50, Cat. #AF1172, R&D Systems, Minneapolis, Minnesota, USA), were employed.

Techniques: Cell Culture, Marker, Light Microscopy, Immunofluorescence, Staining, Fluorescence, Flow Cytometry

Journal: eLife

Article Title: Single-cell profiling reveals the intratumor heterogeneity and immunosuppressive microenvironment in cervical adenocarcinoma

doi: 10.7554/eLife.97335

Figure Lengend Snippet: ( A ) Circle plots showing the interacting networks between epithelial cell sub-clusters and T cell sub-clusters via the pathway of ALCAM by comparing adenocarcinoma (ADC) and squamous cell carcinoma (SCC). ( B ) Circle plots simplified from ( A ) to show the interactions among target cell clusters via ALCAM pathway. ( C ) Circle plots showing the interacting networks between epithelial cell sub-clusters and T cell sub-clusters via the pathway of MHC-II by comparing ADC and SCC. ( D ) Circle plots simplified from ( C ) to show the interactions among target cell clusters via MHC-II pathway. From ( A ) to ( D ), the direction of each arrow shows the regulation from outputting cells to incoming cells. The width of the each line shows the predicted weight and strength of regulation. ( E ) Bubble plot showing the probability of ligand-to-receptor combination of each pathway between two different target sub-clusters of cells by comparing ADC with SCC. ( F ) Circle plots showing Tregs regulate epithelial cells via the TGF-β pathway, which is solely activated in ADC. ( G ) Bubble plot showing the probability of ligand-to-receptor combination of TGF-β pathway between Tregs and epithelial cells by comparing ADC with SCC. The pathways of ADGRE5, CD46, GZMA, and NAMPT are used as negative controls. ( H ) Dual immunofluorescence (IF) staining confirming that in SLC26A3 high regions of CC tissues, more FOXP3 + cells are recruited than in SLC26A3 low regions (left). The numbers of recruited FOXP3 + cell are quantified using histogram plot (right). Three individual samples with ROI were calculated and p<0.01 were marked with **, showing significant difference. ( I ) Multiplexed IF staining confirming the interaction between CD6 (on FOXP3 + cells) and ALCAM (on SLC26A3 high epithelial cells) in the ALCAM pathway. ( J ) Multiplexed IF staining showing that the recruitment of FOXP3 + cells toward SLC26A3 high cells might induce EMT (marked with E-cadherin) and increase the stemness (marked with ALDH1A1) of tumor cells, via TGF-β pathway.

Article Snippet: Antibody , ALCAM polyclonal antibody , Proteintech , Cat# 21972-1-AP , .

Techniques: Immunofluorescence, Staining

Journal: eLife

Article Title: Single-cell profiling reveals the intratumor heterogeneity and immunosuppressive microenvironment in cervical adenocarcinoma

doi: 10.7554/eLife.97335

Figure Lengend Snippet:

Article Snippet: Antibody , ALCAM polyclonal antibody , Proteintech , Cat# 21972-1-AP , .

Techniques: Recombinant