Ephb3

Manufactured by R&D Systems

EphB3 is a protein that functions as a receptor tyrosine kinase. It is involved in the regulation of cellular processes such as cell-cell interaction and cell migration.

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Lab products found in correlation

Ephb2, by R&D Systems (3 mentions) Ephrin b1, by R&D Systems (3 mentions) C myc, by Santa Cruz Biotechnology (2 mentions) Lysozyme, by R&D Systems (1 mentions) Cyclin d1, by Cell Signaling Technology (1 mentions) Chromogranin a, by Immunostar (1 mentions) Cyclin e1, by Abcam (1 mentions) Cyclin a1, by Santa Cruz Biotechnology (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) β catenin, by Cell Signaling Technology (1 mentions) Miz 1, by Santa Cruz Biotechnology (1 mentions) α tubulin, by Merck Group (1 mentions) Myc tag, by Cell Signaling Technology (1 mentions) Vinculin, by Abcam (1 mentions) β actin, by Merck Group (1 mentions) Alexa fluor 647 conjugated phalloidin, by Thermo Fisher Scientific (1 mentions) Sc 5561, by Santa Cruz Biotechnology (1 mentions) A22287, by Thermo Fisher Scientific (1 mentions) Goat anti rabbit 800cw, by LI COR (1 mentions) Hsp70, by BD (1 mentions)

3 protocols using ephb3

Histological Analysis of Mouse Intestinal Tissues

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Small intestine and colon from sacrificed mice were placed immediately into 10% neutral buffered formalin (NBF). Intestines were opened longitudinally, cleaned, and swiss-rolled. Swiss rolls were fixed for ~24 hr in 10% NBF at room temperature (RT). Fixed tissues were embedded in paraffin and stained with H&E. Adenomas were graded by inspection of H&E-stained sections.
For immunohistochemistry (IHC), primary antibodies (Abs) recognized were β-catenin (Cell Signaling Technology, #2677), c-Myc (Santa Cruz, #SC764, N-262), lysozyme (#A0099), ephrinB1 (R&D Systems, clone AF473), EphB2 (R&D Systems, clone AF467), EphB3 (R&D Systems, clone AF432), cleaved caspase-3 (Cell Signaling Technology, #9661), Ki67 (Dako, Tec3 clone), chromogranin A (Immunostar, #20085), p21 (Santa Cruz Biotechnology, clone M19), GFP (Novus, 100–1678), cyclin D1 (Cell Signaling Technology, #2978), cyclin E1 (Abcam, ab7959), cyclin A1 (Santa Cruz Biotechnology, sc-751), and B23 (Sigma, B0556). Anti-LFABP antibody was the kind gift of Dr. Jeffrey Gordon (Washington University, St. Louis).

Dominguez-Brauer C., Hao Z., Elia A.J., Fortin J.M., Nechanitzky R., Brauer P.M., Sheng Y., Mana M.D., Chio I.I., Haight J., Pollett A., Cairns R., Tworzyanski L., Inoue S., Reardon C., Marques A., Silvester J., Cox M.A., Wakeham A., Yilmaz O.H., Sabatini D.M., van Es J.H., Clevers H., Sato T, & Mak T.W. (2016). Mule Regulates the Intestinal Stem Cell Niche via the Wnt Pathway and Targets EphB3 for Proteasomal and Lysosomal Degradation. Cell stem cell, 19(2), 205-216.

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Immunoblotting of Cellular Proteins

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Standard immunoblotting was performed using Abs recognizing HA (Santa Cruz Biotechnology, clone Y-11), Flag (Sigma clone M2), Myc-tag (Cell Signaling Technology, 9B11), vinculin (Abcam, SPM227), Mule (Bethyl Laboratories, A300-486A), ephrinB1 (R&D Systems, clone AF473), EphB2 (R&D clone AF467), EphB3 (R&D Systems, clone AF432), Miz1 (Santa Cruz Biotechnology, sc-22837), c-Myc (Santa Cruz Biotechnology, N262), α-tubulin (Sigma), and β-actin (Sigma). Anti-mouse and anti-rabbit HRP-conjugated secondary Abs were from Thermo Fisher Scientific. Quantification of immunoblots was performed using Image J software (NIH).

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Ephrin-B1 and EphB Receptor Imaging

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Mouse embryos were subjected to immunofluorescence using 2 µg/ml ephrin-B1 (AF473; R&D Systems), 2 µg/ml EphB2 (AF467; R&D Systems), 2 µg/ml EphB3 (AF432; R&D Systems), and 10 µg/ml GFP (ab13970; Abcam) antibodies. The following secondary antibodies were used: Alexa Fluor 488–conjugated donkey anti–chicken IgG (1:1,000, 703-545-155; Jackson ImmunoResearch Laboratories, Inc.) and Cy3-conjugated donkey anti–goat IgG (1:400; 705-165-003; Jackson ImmunoResearch Laboratories, Inc.). To label F-actin, Alexa Fluor 647–conjugated phalloidin (1:40; A22287; Thermo Fisher Scientific) was used. For Western blotting, the following primary antibodies were used: rabbit anti-ROCK1 (1:500; sc-5560; Santa Cruz Biotechnology, Inc.), ROCK2 (1:500; sc-5561; Santa Cruz Biotechnology, Inc.), and HSP70 (1:1,000; 610607; BD). The following IRDye secondary antibodies were used: goat anti–mouse 680RD (1:5,000; 925–68070; LI-COR Biosciences) and goat anti–rabbit 800CW (1:5,000; 926–32211; LI-COR Biosciences).

O’Neill A.K., Kindberg A.A., Niethamer T.K., Larson A.R., Ho H.Y., Greenberg M.E, & Bush J.O. (2016). Unidirectional Eph/ephrin signaling creates a cortical actomyosin differential to drive cell segregation. The Journal of Cell Biology, 215(2), 217-229.

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