- 32-4000 - Invitrogen Antibodies CDH3 antibody

Submitted references Biphasic Squamoid Alveolar Renal Cell Carcinoma: A Distinctive Subtype of Papillary Renal Cell Carcinoma?

Hes O, Condom Mundo E, Peckova K, Lopez JI, Martinek P, Vanecek T, Falconieri G, Agaimy A, Davidson W, Petersson F, Bulimbasic S, Damjanov I, Jimeno M, Ulamec M, Podhola M, Sperga M, Pane Foix M, Shelekhova K, Kalusova K, Hora M, Rotterova P, Daum O, Pivovarcikova K, Michal M
The American journal of surgical pathology 2016 May;40(5):664-75

HIV-1 Nef and KSHV oncogene K1 synergistically promote angiogenesis by inducing cellular miR-718 to regulate the PTEN/AKT/mTOR signaling pathway.

Xue M, Yao S, Hu M, Li W, Hao T, Zhou F, Zhu X, Lu H, Qin D, Yan Q, Zhu J, Gao SJ, Lu C
Nucleic acids research 2014 Sep;42(15):9862-79

Novel three-dimensional in vitro models of ovarian endometriosis.

Brueggmann D, Templeman C, Starzinski-Powitz A, Rao NP, Gayther SA, Lawrenson K
Journal of ovarian research 2014 Feb 6;7:17

A three-dimensional microenvironment alters protein expression and chemosensitivity of epithelial ovarian cancer cells in vitro.

Lee JM, Mhawech-Fauceglia P, Lee N, Parsanian LC, Lin YG, Gayther SA, Lawrenson K
Laboratory investigation; a journal of technical methods and pathology 2013 May;93(5):528-42

P-cadherin is a direct PAX3-FOXO1A target involved in alveolar rhabdomyosarcoma aggressiveness.

Thuault S, Hayashi S, Lagirand-Cantaloube J, Plutoni C, Comunale F, Delattre O, Relaix F, Gauthier-Rouvière C
Oncogene 2013 Apr 11;32(15):1876-87

Sporadic hybrid oncocytic/chromophobe tumor of the kidney: a clinicopathologic, histomorphologic, immunohistochemical, ultrastructural, and molecular cytogenetic study of 14 cases.

Petersson F, Gatalica Z, Grossmann P, Perez Montiel MD, Alvarado Cabrero I, Bulimbasic S, Swatek A, Straka L, Tichy T, Hora M, Kuroda N, Legendre B, Michal M, Hes O
Virchows Archiv : an international journal of pathology 2010 Apr;456(4):355-65

JNK signaling pathway is required for bFGF-mediated surface cadherin downregulation on HUVEC.

Wu JC, Yan HC, Chen WT, Chen WH, Wang CJ, Chi YC, Kao WY
Experimental cell research 2008 Feb 1;314(3):421-9

Differences in invasive capacity of endometrial cancer cell lines expressing different progesterone receptor isotypes: possible involvement of cadherins.

Hanekamp EE, Gielen SC, De Ruiter PE, Chadha-Ajwani S, Huikeshoven FJ, Burger CW, Grootegoed JA, Blok LJ
Journal of the Society for Gynecologic Investigation 2005 May;12(4):278-84

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of P-Cadherin was performed by loading 20 µg of PC-3 (lane1), MCF7 (lane2) and A431 (lane3) using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800). Proteins were transferred to a PVDF membrane and blocked with 5 % skim milk for 1 hour at room temperature. P-Cadherin was detected at ~110 kDa using P-Cadherin Mouse Monoclonal Antibody (Product # 32-4000) at 1-3 µg/mL dilution in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Mouse IgG - HRP Secondary Antibody (Product # 62-6520) at 1:4000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of total Cadherin and P-Cadherin was performed by loading and 2 µL SeeBlue® Plus2 Prestained Protein Ladder (Product # LC5925), 50 µg of MDCK cell lysates per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 1% BSA/TBST for at least 1 hour at room temperature. Total cadherin was detected using a rabbit antibody (Product # 71-7100) and P-Cadherin was detected using a mouse antibody (Product # 32-4000), both at a concentration of 1 µg/mL in blocking buffer overnight at 4°C on a rocking platform. The blot was then incubated with a goat anti-rabbit IgG-Alexa fluor 790 secondary antibody (Product # A11369) and a goat anti-mouse IgG-Alexa Fluor 680 secondary antibody (Product # A-21058) at a dilution of 1:10,000 for at least 1 hour. Fluorescent detection was performed using the Odyssey® CLx imaging system (Li-cor Biosciences). Images is generated by Joell Solan in Paul Lampe Lab at Fred Hutchinson Cancer Research Center.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of P-cadherin was achieved by transfecting MCF7 cells with P-cadherin specific siRNAs (Silencer® select Product # s533726, s533725). Western blot analysis (Fig. a) was performed using whole cell extracts from the P-cadherin knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with P-cadherin Monoclonal Antibody (12H6) (Product # 32-4000, 1µg/mL) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Reduction in signal upon siRNA mediated knock down confirms that antibody is specific to P-cadherin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of P-Cadherin was performed on cryosections of human skin tissue. Tissues were blocked in 10% normal goat serum in 1X PBS containing 0.1% Triton X-100 (1X PBS-T) for 1 hour at room temperature (RT). The tissues were labeled with a P-Cadherin monoclonal antibody (clone 12H6, red, Product # 32-4000) diluted 1:50 in 3% normal goat serum in 1X PBS-T for 1 hour at RT, followed by detection with a Goat anti-Mouse IgG1, Alexa Fluor 568 secondary antibody (Product # A-21124) diluted 1:2000 in 3% normal goat serum in 1X PBS-T for 1 hour at RT. Nuclei (blue) were stained with DAPI, included in ProLong Gold Anti-Fade Mountant (Product # P36931). Images were taken on an inverted microscope at 40X magnification. Data courtesy of Dr. Jiyoon Lee at Indiana University School of Medicine.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7. miR-718 mediates K1- and Nef-induced angiogenesis both in vitro and in vivo . ( A ) Matrigel assay analysis of microtubule formation. Tube formation assay was performed with HUVECs transduced with K1, incubated with soluble Nef protein for 72 h or both and further transfected with negative control nucleotide of miRNA (Neg. Ctrl.; top) or inhibitor of miR-718 (miR-718 inhibitor; bottom) for 48 h, respectively. The photographs of microtubules were captured at 16 h post seeding (original magnification, x100). ( B ) Quantification of results in (A). The results represent the mean +- SD from three independent experiments ( n = 3), each experiment containing six technical replicates. ( C ) Inhibition of miR-718 suppressed the enhanced effect of Nef on K1-induced angiogenesis. HUVECs transduced with K1, incubated with soluble Nef protein for 72 h or both were transfected with negative control nucleotide of miRNA (Neg. Ctrl.; top) or inhibitor of miR-718 (miR-718 inhibitor; bottom) for 48 h. The collected cells were mixed with Matrigel and subsequently implanted onto the CAM. Representative photographs of angiogenesis on the CAM are shown. ( D ) Quantification of results in (C). The number of blood vessels is expressed as the mean +- SD from three independent experiments ( n = 3), each experiment containing six technical replicates. ( E ) Western blot analysis of total PTEN and phosphorylation levels of AKT and mTOR. The tumor tissues from CAM that treated as in (C) were co

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 8. miR-718 mediates K1- and Nef-induced tumorigenesis in nude mice. ( A ) Inhibition of miR-718 abrogates the enhanced effect of Nef on K1-induced tumorigenesis. EA.hy926 cells transduced with K1, Nef or both were infected with control virus (pCDH) or miR-718 sponge for 72 h and further re-suspended in serum-free medium. As detailed in the 'Materials and Methods' section, the treated cells were injected (s.c.) into nude mice. The sizes of tumors from nude mice were determined by two-dimensional caliper measurements. Data represent mean +- SD, each group with five tumors ( n = 5). Two independent experiments were performed and similar results were obtained. ** and *** indicate P < 0.01 and P < 0.001 by Student's t -test, respectively. ( B ) Tumor-bearing mice were killed at day 56 after injections, and tumors were removed and pictures were taken. ( C ) Inhibition of miR-718 abrogates the enhanced effect of Nef on K1-induced tumorigenesis. The tumors from nude mice treated as in (A) were removed and weighed. Scatter plots represent the weights of independent tumors from different groups. Data represent mean +- SD, each group with five tumors ( n = 5). Two independent experiments were performed and similar results were obtained. ( D ) Inhibition of miR-718 increased total PTEN level and suppressed the enhanced phosphorylation of AKT and mTOR by K1 and Nef. The tumor tissues from nude mice treated as in (A) were removed, and the expression of total PTEN, pho

32-4000

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