Heca 452

Manufactured by BioLegend

HECA-452 is a mouse monoclonal antibody that recognizes the human endothelial cell antigen-452. It is designed for use in flow cytometry applications.

Automatically generated - may contain errors

Lab products found in correlation

Goat anti rat igg hrp, by Southern Biotech (2 mentions) Kpl 1, by BD (2 mentions) Hrp conjugated secondary antibody, by Southern Biotech (1 mentions) Rat anti mouse cd62e mab, by R&D Systems (1 mentions) Lumi light western blotting substrate, by Roche (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) Cytomics fc500 mpl flow cytometer, by Beckman Coulter (1 mentions) Protein g agarose, by Thermo Fisher Scientific (1 mentions) Reducing sds page gels, by Bio-Rad (1 mentions) Facscanto 2, by BD (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions)

3 protocols using heca 452

Western Blotting Analysis of Leukocyte Adhesion Molecules

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein samples were boiled in reducing Laemmli loading buffer (Boston BioProducts) and then resolved on 7.5% SDS-PAGE electrophoresis gels (Bio-Rad). Resolved proteins were transferred to PVDF membranes (Bio-Rad) and blocked with 10% milk and 0.1% Tween20 in TBS. Western blots were probed with primary antibodies (1 μg/mL), followed by incubation with HRP-conjugated secondary antibodies (Southern Biotech). Expression of sLe^X, CD44, PSGL-1, CD43, MPO and L-selectin were assessed using the mAbs HECA-452 (Biolegend), 2C5 (R&D Systems), KPL-1 (BD Biosciences), 1G10 (BD Biosciences), 2C7 (Abcam) and LAM1-116 (Santa Cruz Biotechnology), respectively. For E-Ig blotting, membranes were incubated with E-Ig (1 μg/mL) suspended in TBS 0.1% Tween20 containing 2 mM CaCl_2, followed by incubation with rat anti-mouse CD62E mAb (R&D Systems) and then goat anti-rat IgG-HRP (Southern Biotech). Antigens were detected by chemiluminescence using Lumi-Light Western blotting substrate (Roche).

Silva M., Fung R.K., Donnelly C.B., Videira P.A, & Sackstein R. (2017). Cell–specific Variation in E-selectin Ligand Expression Among Human Peripheral Blood Mononuclear Cells: Implications for Immunosurveillance and Pathobiology. Journal of immunology (Baltimore, Md. : 1950), 198(9), 3576-3587.

+ Open protocol

+ Expand

Phenotypic Characterization of hMSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

hMSCs were incubated with a panel of primary antibodies chosen based on the known profile of BM-hMSC populations [3 ,4 (link)]. Flow cytometry analysis was performed using the following commercially available mAbs together with isotype-matched controls: FITC-conjugated anti-human CD13 and CD36, and PE-conjugated anti-human CD34, CD105, CD146, and CD166 (all from Beckman Coulter); FITC-conjugated mAb HECA452 (Biolegend) and anti-human CD62E (R&D Systems); PE-conjugated anti-human CD44, CD45, CD49d, CD73 and CD90, and PECy5-conjugated anti- human CD29 and CD106 (all from BD Biosciences). Data were collected using a Cytomics FC 500 MPL flow cytometer (Beckman Coulter) and analyzed with FlowJo version 10.0.6 (TreeStar).

Pachón-Peña G., Donnelly C., Ruiz-Cañada C., Katz A., Fernández-Veledo S., Vendrell J, & Sackstein R. (2017). A Glycovariant of Human CD44 is Characteristically Expressed on Human Mesenchymal Stem Cells. Stem cells (Dayton, Ohio), 35(4), 1080-1092.

+ Open protocol

+ Expand

Flow Cytometry and Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Flow cytometry was performed on a FACS Canto II (BD Biosciences) and analyzed using FlowJo Software (Treestar). Cells were stained with antibodies to CD3, CD4 (OKT4), CXCR4 (12G5), VLA-4 (9F10) and CD15s (CSLEX1) from BD biosciences, and HECA452, CD45 (2D1), CD44 (BJ14), CD162 (KPL-1), CD43 (CD43-10G7), and FoxP3 (206D) from Biolegend. Staining with mouse E-selectin Ig chimera (RnD Systems) was detected with either anti-His FITC (Bethyl Laboratories) for flow cytometry or secondary rat anti-mouse CD62E (BBIG-E4, RnD Systems) followed by goat anti-rat IgG HRP (Southern Biotech) for western analyses.
Lysates were made from cells by sonication/vortexing in buffer containing 50 mM Tris, 150 mM NaCl, 20 ug/ml phenylmethanesulfonyl fluoride (PMSF), 0.2% NaN₃, protease inhibitor cocktail (Roche), 2% NP40, and 0.2% SDS. Precleared lysates were incubated antibodies to CD43 (1G10, L60, BD Biosciences; 20819, Santa Cruz Biotechnology), PSGL-1 (KPL-1, BD Biosciences; 20929, Santa Cruz Biotechnology), or CD44 (515, BD Biosciences; 2C5, RnD systems). Protein was immunoprecipitated with Protein-G agarose (Life Technologies). Western Blots were run with Reducing SDS-PAGE gels (Bio-Rad).

Donnelly C., Dykstra B., Mondal N., Huang J., Kaskow B.J., Griffin R., Sackstein R, & Baecher-Allan C. (2018). Optimizing human Treg immunotherapy by Treg subset selection and E-selectin ligand expression. Scientific Reports, 8, 420.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!