Histological and immunohistochemical analyses were performed as previously described2 (link). The following antibodies were used: Cdh1 (rat, Novex), Cortactin (rabbit, Abcam), GFP (goat, Abcam), GFP (mouse, Roche), ItgA2 (rabbit, Abcam), ItgA6 (rabbit, Abcam), ItgB1 (rabbit, Abcam), Ki67 (rat, Biolegend), Krt19 TROMA-III (rat, DSHB), MLC2 pSer19 (rabbit, NEB), Myosin (rabbit, Abcam), Nestin (mouse, BD transduction), Pdgfrβ (rabbit, NEB), PTK2 pTyr397 (rabbit, ThermoFisher), SMA (mouse, Agilent), SMA (mouse, Sigma), Tomato (rabbit, Rockland), Tomato (goat, Biorbyt), Vimentin (rabbit, NEB), Vinculin (mouse, Sigma-Aldrich). DBA-rhodamine and DBA-FITC were from Vectorlabs. F-actin was stained with Phalloidin-TRITC (Sigma-Aldrich) and nuclei with DAPI (Sigma-Aldrich). F-actin staining of LSL-KrasG12D; p53 F/F; Pdx1-Cre pancreata and wildtype littermate control pancreata was performed on cryosections. Samples were embedded fresh in OCT medium and after sectioning fixed in 5% NBF for 10 min. Slides were washed in 0.2% Triton X-100 in PBS for 10 min and incubated in FLASH blocking buffer for 30 min. Staining reagent incubations were performed as above. Fluorescent stainings were imaged on a Zeiss LSM 780 confocal microscope. Chromogenic DAB stainings were imaged on a Zeiss Axio Scan Z1 Slide Scanner.
Itga6
Manufactured by Abcam
Sourced in United Kingdom
ITGA6 is a protein that plays a role in cell-cell and cell-matrix interactions. It is a member of the integrin alpha chain family and is expressed on the cell surface.
Automatically generated - may contain errors
Lab products found in correlation
Cortactin, by Abcam (1 mentions)
Itga2, by Abcam (1 mentions)
Dba fitc, by Vector Laboratories (1 mentions)
Dba rhodamine, by Vector Laboratories (1 mentions)
Axio scan z1 slide scanner, by Zeiss (1 mentions)
Phalloidin tritc, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Vinculin, by Merck Group (1 mentions)
Nestin, by BD (1 mentions)
Lsm 780 confocal microscope, by Zeiss (1 mentions)
Itgb1, by Abcam (1 mentions)
Enhanced chemiluminescence detection system, by Merck Group (1 mentions)
Anti β actin peroxidase antibody, by Merck Group (1 mentions)
Protease inhibitor, by Merck Group (1 mentions)
Quantity one 4, by Bio-Rad (1 mentions)
β actin, by Merck Group (1 mentions)
Krt14, by BioLegend (1 mentions)
Krt18, by Abcam (1 mentions)
Phospho p44 42 mapk erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
3 protocols using itga6
1
Comprehensive Tissue Analysis Protocol
2
Quantitative Protein Expression Analysis
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Total protein lysates were extracted with lysis buffer (1% Triton, 50 nmol/L Tris, 15 mmol/L NaCl) supplemented with protease inhibitors (Sigma–Aldrich, St. Louis, MO, USA). The following primary antibodies were used:: ITGA6 (recombinant rabbit monoclonal antibody, clone: EPR18124, catalog number: ab181551, Abcam), E2F1 (mouse polyclonal antibody, clone: C-20, catalog number: sc-193, Santa Cruz), YY1 (mouse monoclonal antibody, clone: 2E11C5, catalog number: 66281-1-Ig, Proteintech, Manchester, United Kingdom), E2F2 (mouse monoclonal antibody, clone: TFE-25, catalog number: sc-9967, Santa Cruz Biothecnology, Dallas, TX, USA), β-actin (mouse monoclonal Anti-β-actin−Peroxidase antibody, clone: AC-15, catalog number: A3854, Sigma–Aldrich, St. Louis, MO, USA) and vinculin (mouse monoclonal antibody, clone: hVIN-1, catalog number: V9131, Sigma–Aldrich, St. Louis, MO, USA). Proteins were visualized by enhanced chemiluminescence detection system (Sigma–Aldrich, St. Louis, MO, USA). Quantification was performed by Quantity One 4.6.6 software (Bio-Rad, Hercules, California, USA).
3
Immunostaining Protocols for Cell and Tissue Analysis
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Primary antibodies used for immunocytochemistry were 1:1000 KRT14 (BioLegend, 905301), 1:500 KRT18 (abcam, ab24561), 1:500 KRT8/18 (Invitrogen, MA5–14088), 1:300 active ITGβ1 (Sigma Aldrich, MAB2079Z), and 1:200 phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology, 9101S). Primary antibodies used for immunohistochemistry of FFPE tissues were 1:50 COL6 (Rockland, 600–401-108–0.1) and 1:500 ITGA6 (abcam, ab181551). Secondary antibodies used for both methods, all diluted 1:1,000, were Alexa Fluor 488 Goat anti-Rabbit IgG Highly Cross-Adsorbed (Invitrogen, A-11034), Alexa Fluor 647 Goat anti-Mouse IgG Cross-Adsorbed (Invitrogen, A-21235), and Alexa Fluor™ 546 Goat anti-Rabbit IgG Cross-Adsorbed (Invitrogen, A-11010).
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