Dlk 1

Manufactured by Abcam

Sourced in China

DLK-1 is a recombinant protein that functions as a protein kinase. It is involved in the regulation of cell proliferation and differentiation.

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Lab products found in correlation

6 protocols using dlk 1

Comprehensive Immunohistochemical Analysis of Tissue Samples

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The following antibodies were used: N-cadherin (clone GC4, Sigma-Aldrich C3865, 1:100 dilution), FSP1 (Abcam ab58597, 1:100 dilution), PDGFRα (R&D Systems AF1062, 1:50 dilution), Collagen I (Rockland 600–401–103–0.1, 1:150 dilution), Collagen III (Abcam ab7778, 1:150 dilution), Fibronectin (Abcam ab23750, 1:200 dilution), α-SMA (Abcam ab5694, 1:100 dilution), CD31 (Novus Biologicals NB100-2284, 1:500 dilution), EpCAM (Abcam ab92382, 1:500 dilution), Lyve-1 (Abcam ab14917, 1:500 dilution), Myf-5 (clone C-20, Santa Cruz sc-302, 1:500 dilution), CD45 (clone IBL-3/16, Abcam ab23910, 1:500 dilution), FABP4 (clone EPR3579, Abcam ab92501, 1:500 dilution), F4/80 (Abcam ab90247, 1:500 dilution), Cytokeratin 14 (clone EPR17350, Abcam ab181595, 1:200 dilution), Integrin α_v (clone EPR16800, Abcam ab179475, 1:500 dilution), α1-catenin (clone EP1793Y, Abcam ab51032, 1:500 dilution), decorin (Abcam ab175404, 1:500 dilution), DLK-1 (Abcam ab21682, 1:200 dilution) and Ki67 (clone SP6, Abcam ab16667, 1:500 dilution). Fluorophore-conjugated secondary antibodies were purchased from Thermo Fisher Scientific (1:500 dilution). Exherin (ADH-1) was from TargetMol (T2637).

Jiang D., Christ S., Correa-Gallegos D., Ramesh P., Kalgudde Gopal S., Wannemacher J., Mayr C.H., Lupperger V., Yu Q., Ye H., Mück-Häusl M., Rajendran V., Wan L., Liu J., Mirastschijski U., Volz T., Marr C., Schiller H.B, & Rinkevich Y. (2020). Injury triggers fascia fibroblast collective cell migration to drive scar formation through N-cadherin. Nature Communications, 11, 5653.

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Histological Analysis of Liver Fibrosis

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Formalin-fixed samples were embedded in paraffin, cut into 5 μm-thick sections and stained with hematoxylin-eosin (H-E), Masson’s trichrome and Sirius red according to standard procedures. Trichrome stained sections were analyzed to score the amount of fibrosis according to the method of Ishak.^{25 (link)} The collagen proportional area (CPA) was morphometrically quantified on Sirius red stained sections with image processing software (ImageJ, NIH). Additional sections were stained with an antibodies specific for receptor for advanced glycation end products (RAGE), delta-like homologue 1 (DLK-1) (both from Abcam, Cambridge, MA) and OV-6 (R&D Systems, Minneapolis, MN). All slides were reviewed blindly by the same liver pathologist.

DePeralta D.K., Wei L., Ghoshal S., Schmidt B., Lauwers G.Y., Lanuti M., Chung R.T., Tanabe K.K, & Fuchs B.C. (2016). Metformin Prevents Hepatocellular Carcinoma Development by Suppressing Hepatic Progenitor Cell Activation in a Rat Model of Cirrhosis. Cancer, 122(8), 1216-1227.

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Dlk1 Overexpression Protein Analysis

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The total protein content was extracted from cells of each cell line by using RIPA lysis buffer and then determined by using the BCA method. The whole protein of each sample was separated using 10% SDS-PAGE and transferred onto PVDF membranes for blotting. The antibodies used in this study were Gapdh (#60004-1-Ig Proteintech, Wuhan, China), Dlk1 (#ab210471, Abcam, Cambridge, UK), Notch1 (#4ab094535, 4A BIOTECH, Beijing, China), Rhoc (#4ab090482, 4A BIOTECH, Beijing, China), Erk1/2 (#BF8004, Affinity Biosciences, Changzhou, China), and p-Erk1/2 (#AF1015, Affinity Biosciences, Changzhou, China). The full length of Dlk1 cDNA was cloned to pcDNA3.1 to overexpress Dlk1.

Han X., He H., Shao L., Cui S., Yu H., Zhang X, & Wu Q. (2022). Deletion of Meg8-DMR Enhances Migration and Invasion of MLTC-1 Depending on the CTCF Binding Sites. International Journal of Molecular Sciences, 23(15), 8828.

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Protein Expression Analysis Protocol

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Anti-Prmt5 (Millipore), -β-catenin (Cell Signaling), Histone H3 (Millipore), -Dlk-1 (Abcam) and -Tubulin antibodies were used.

Paul C., Sardet C, & Fabbrizio E. (2015). The Wnt-target gene Dlk-1 is regulated by the Prmt5-associated factor Copr5 during adipogenic conversion. Biology Open, 4(3), 312-316.

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DLK-1 Expression Modulation in HUVECs

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For DLK-1 experiments, HUVECs seeded in 6-well plates were incubated with nanocomplexes at a 50:1 ratio containing 1.0µg of negative siRNA (Allstars, Qiagen) or miR-126–5p for 48 hours. The cellular proteins were extracted and lysates were processed as described above. Blots were probed for α-tubulin (1:1000; Cell Signaling), or DLK-1 (1:1000; Abcam) overnight at 4°C and IRDye secondary antibodies (1:10000) for 1 hour. Blots were analyzed using an Odyssey Licor Imaging System (Li-COR).

Jones Buie J.N., Zhou Y., Goodwin A.J., Cook J.A., Vournakis J., Demcheva M., Broome A.M., Dixit S., Halushka P.V, & Fan H. (2019). Application of deacetylated-poly-N-acetyl glucosamine nanoparticles for the delivery of miR-126 for the treatment of cecal-ligation puncture induced sepsis. Inflammation, 42(1), 170-184.

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Immunohistochemical Analysis of Fetal Liver Markers

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Tissues were either fixed overnight and embedded in paraffin. Sections were cut 4–5 mm thick. Paraffin sections were deparaffinized, dehydrated, and we performed antigen retrieval by steaming slides in sodium citrate buffer for 30 min. Sections were blocked in the blocking serum buffer (5% serum in PBS + 0.5% Triton X-100) for 30 min. Primary antibodies were diluted in blocking buffer and incubated on tissue sections overnight at 4 °C. Slides were washed and incubated in secondary antibody in blocking buffer for 2 h at room temperature. Slides were washed and mounted using Fluormount-G and observed under a microscope. Antibody information: FOXA2 (Cell Signaling, #3143); DLK1 (Abcam, ab21682); HNF4A (LSBio, LS-C413074); LZTS1 (Bioss, bs-5705R).

Mu T., Xu L., Zhong Y., Liu X., Zhao Z., Huang C., Lan X., Lufei C., Zhou Y., Su Y., Xu L., Jiang M., Zhou H., Lin X., Wu L., Peng S., Liu S., Brix S., Dean M., Dunn N.R., Zaret K.S., Fu X.Y, & Hou Y. (2020). Embryonic liver developmental trajectory revealed by single-cell RNA sequencing in the Foxa2eGFP mouse. Communications Biology, 3, 642.

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