36-6200

Mansouri R, Jouan Y, Hay E, Blin-Wakkach C, Frain M, Ostertag A, Le Henaff C, Marty C, Geoffroy V, Marie PJ, Cohen-Solal M, Modrowski D
Cell death & disease 2017 Jun 29;8(6):e2902

Syndecan-2 is a key regulator of transforming growth factor beta 2/Smad2-mediated adhesion in fibrosarcoma cells.

Mytilinaiou M, Bano A, Nikitovic D, Berdiaki A, Voudouri K, Kalogeraki A, Karamanos NK, Tzanakakis GN
IUBMB life 2013 Feb;65(2):134-43

Differences in heparan sulfate production in cervical fibroblast cultures from women undergoing term and preterm delivery.

Akerud A, Dubicke A, Sennstrom M, Ekman-Ordeberg G, Malmstrom A
Acta obstetricia et gynecologica Scandinavica 2008;87(11):1220-8

CCN2 (connective tissue growth factor) promotes fibroblast adhesion to fibronectin.

Chen Y, Abraham DJ, Shi-Wen X, Pearson JD, Black CM, Lyons KM, Leask A
Molecular biology of the cell 2004 Dec;15(12):5635-46

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of (A) A549, (B) HepG2, (C) HT-20, (D) K-562, (E) TF-1, and (F) U2-OS cell lysates using Rb anti-Syndecan-2 (Product # 36-6200).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Syndecan-2 was performed by loading 20 µg of K562 (lane1) HT-29 (lane2), Hep G2 (lane3), A549 (lane4) and DU 145 (lane5) cell lysate using Novex®NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® 2 Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk at 4°C overnight. Syndecan-2 was detected at ~ 22 and 37 kDa using Syndecan-2 Rabbit Polyclonal Antibody (Product # 36-6200) at 1-2 µg/mL in 5% skim milk for 3 hour at room temperature on a rocking platform. Goat Anti-Rabbit IgG - HRP Secondary Antibody (G21234) at 1:5000 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: Immunofluorescence analysis of Syndecan-2 was done on 90% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Syndecan-2 Rabbit polyclonal Antibody (Product # 36-6200) at 2 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing junctional localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis ofSyndecan-2 showing staining in the cytoplasm of paraffin-embedded human hepatocellular carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Syndecan-2 polyclonal antibody (Product # 36-6200) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Syndecan-2 was done on HeLa cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Syndecan-2 Rabbit Polyclonal Antibody (366200, red histogram) or with rabbit isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor¨ 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Syndecan-2 overexpression enriched bone surface with heparan sulfate glycosaminoglycans. ( a ) Immunohistological analysis of syndecan-2 in paraffin-embedded vertebra from 2-month-old WT ( a and b ) or ColI-Synd2 ( c and d ) mice. ( b ) Flow cytometry analysis of syndecan-2 expression. Marrow cells from 6-week-old mice were flushed from long bones and labeled with surface markers to select osteoblast precursors (CD51 + Sca-1 + cells) or osteoblasts (CD51 + Sca-1 - cells). Fluorescence intensity versus structural parameter of the cells (FSC-H) was plotted. ( c ) The median fluorescence intensity (MFI) of syndecan-2 labeling was recorded in different cell populations from WT or ColI-Synd2 mice. Data are mean+-S.E.M. ( n =7 WT and three ColI-Synd2 mice). ( d ) The MFI of heparan sulfate labeling was recorded in CD51 + Sca-1 - cells from the marrow of WT or ColI-Synd2 mice. Data are mean+-S.E.M. ( n =4 WT and four ColI-Synd2 mice). ( e ) Immunohistological analysis of heparan sulfate chains in paraffin-embedded vertebra from 2-month-old WT or ColI-Synd2 mice. The photos presented are representative views of syndecan-2 or heparan sulfate staining. Arrows indicate syndecan-2- or heparan sulfate-expressing cells. Dashed lines indicate bone surfaces. *Indicates significant difference between WT and ColI-Synd2 ( P