rabbit anti agr2 monoclonal antibody  (Cell Signaling Technology Inc)

Journal: Cell Death & Disease

Article Title: Vitamin D opposes multilineage cell differentiation induced by Notch inhibition and BMP4 pathway activation in human colon organoids

doi: 10.1038/s41419-024-06680-z

Figure Lengend Snippet: A Light microscopy images of colon normal organoids cultured in PROL, DIFF, or B + D media for 72 h (patient #110). Scale bar, 500 μm. B RT-qPCR analysis of the RNA levels of stemness, enterocytic and mucosecretory genes in organoids from patients #47, #86, #110, #130 for 48 h and #47, #110, #130, #159 for 72 h cultured in PROL (black), DIFF (red) or B + D (green) media. * P < 0.05, ** P < 0.01, *** P < 0.001. C Immunofluorescence analysis of the expression of enterocytic and mucosecretory protein markers (green) in organoids cultured for 48 h in PROL or B + D media (patient #158). Scale bars, 30 µm. Box-plots show the quantification of the fluorescence intensity (Log 2 MGV) (patients #92, #158 and #166; the number of organoids analyzed was 55 for KLF4, 80 for KRT19, 83 for AGR2 and 79 for TFF2). ** P < 0.01, *** P < 0.001. MGV: mean gray value.

Article Snippet: Non-specific sites were blocked by incubation in PBS 2% BSA at RT for 1 h. Organoids were then incubated overnight at 4 °C with the primary antibodies diluted in PBS 1% BSA: rabbit monoclonal-AGR2 (1/100, Cell Signalling, #13062), mouse monoclonal-E-cadherin (1/100, BD Biosciences, NJ, USA, #610182), rabbit polyclonal-KLF4 (1/500, Gene Tex, CA, USA, #GTX101509), mouse monoclonal-KRT19 (1/10, Progen, Germany, #61010), rabbit polyclonal-TFF2 (1/100, Proteintech, #13681-1-AP).

Techniques: Light Microscopy, Cell Culture, Quantitative RT-PCR, Immunofluorescence, Expressing, Fluorescence

Journal: Cell Death & Disease

Article Title: Vitamin D opposes multilineage cell differentiation induced by Notch inhibition and BMP4 pathway activation in human colon organoids

doi: 10.1038/s41419-024-06680-z

Figure Lengend Snippet: A Light microscopy images of organoids (patient #110) cultured for 72 h in DIFF, BMP4, DBZ or B + D media in the absence (vehicle) or presence of calcitriol (100 nM). Scale bar, 500 μm. B RT-qPCR analysis of the RNA levels of enterocytic ( KRT20 , CA1 , KLF4 and FABP2 ) and mucosecretory ( AGR2 , TFF2 and TFF3 ) genes in organoids from patients #47, #86, #110, #130 for 48 h and #47, #110, #130, #159 for 72 h cultured in DIFF (red), BMP4 (blue), DBZ (orange) or B + D (green) media in the absence (vehicle) or presence of calcitriol (100 nM). * P < 0.05, ** P < 0.01, *** P < 0.001. C RT-qPCR analysis of the RNA levels of stemness genes in organoids from the same patients and in the same conditions as in B. D Western blot analysis and quantification of the effect of calcitriol on the expression of enterocytic (CA1) and mucosecretory (TFF2) marker proteins in organoids from four patients incubated in PROL or B + D media in the absence (vehicle) or presence of calcitriol (100 nM) for 48 h. The stemness PTK7 protein was used as control of differentiation and GAPDH as loading control. * P < 0.05. E Immunofluorescence analysis of the expression of enterocytic and mucosecretory protein markers in organoids (patient #158) cultured for 48 h in PROL or B + D media in the absence (vehicle) or presence of calcitriol (100 nM). Scale bars, 30 µm. Box-plots show the quantification of fluorescence intensity (Log 2 MGV) (patients #92, #158 and #166; the number of organoids analyzed was 68 for KLF4, 104 for KRT19, 110 for AGR2 and 79 for TFF2). ** P < 0.01, *** P < 0.001. F Western blot analysis and quantification of the level of phospho(P)-SMAD1/5/8 in organoid cultures that were incubated for 24 h with calcitriol (100 nM) or vehicle before incubation in DIFF medium or BMP4 medium (50 ng/mL) during the indicated times. MGV: mean gray value.

Article Snippet: Non-specific sites were blocked by incubation in PBS 2% BSA at RT for 1 h. Organoids were then incubated overnight at 4 °C with the primary antibodies diluted in PBS 1% BSA: rabbit monoclonal-AGR2 (1/100, Cell Signalling, #13062), mouse monoclonal-E-cadherin (1/100, BD Biosciences, NJ, USA, #610182), rabbit polyclonal-KLF4 (1/500, Gene Tex, CA, USA, #GTX101509), mouse monoclonal-KRT19 (1/10, Progen, Germany, #61010), rabbit polyclonal-TFF2 (1/100, Proteintech, #13681-1-AP).

Techniques: Light Microscopy, Cell Culture, Quantitative RT-PCR, Western Blot, Expressing, Marker, Incubation, Control, Immunofluorescence, Fluorescence

Journal: Cell reports

Article Title: The Lineage Determining Factor GRHL2 Collaborates with FOXA1 to Establish a Targetable Pathway in Endocrine Therapy-Resistant Breast Cancer

doi: 10.1016/j.celrep.2019.09.032

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Monoclonal Anti-AGR2 , Novus Bio , Cat#NBP1–40630; RRID:AB_2305344.

Techniques: Microarray, Recombinant, cDNA Synthesis, SYBR Green Assay, Sample Prep, ChIP-sequencing, Negative Control, Library Amplification, Software, Cell Culture, Membrane, Magnetic Beads, DNA Purification, Protease Inhibitor