Epha2

Manufactured by R&D Systems

EphA2 is a recombinant human protein that represents the extracellular domain of the Ephrin type-A receptor 2. EphA2 is a member of the Eph receptor tyrosine kinase family and plays a role in cell-cell adhesion and migration.

Automatically generated - may contain errors

Lab products found in correlation

Fcs express software, by De Novo Software (1 mentions) Epha3, by R&D Systems (1 mentions) Ephb2, by R&D Systems (1 mentions) Versene, by Thermo Fisher Scientific (1 mentions) F actin, by Thermo Fisher Scientific (1 mentions) Alexa448 conjugated phalloidin, by Thermo Fisher Scientific (1 mentions) P myosin, by Cell Signaling Technology (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) β catenin, by BD (1 mentions) N cadherin, by BD (1 mentions) Cd112, by R&D Systems (1 mentions) Cd155, by R&D Systems (1 mentions) Phosphate buffered saline (pbs), by Merck Group (1 mentions) Facscanto, by BD (1 mentions)

3 protocols using epha2

Antibody Staining and Flow Cytometric Analysis

Cited 1 time

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

Cells were detached with Versene (Invitrogen) counted and resuspended in 100 µL phosphate buffered saline (PBS) containing 1% bovine serum albumin (BSA). Cells were blocked with PBS/1% BSA for 1 h on ice. Two micrograms of QUAD 3.0, EphA3 antibody (house made),^{9 (link)} EphA2, EphB2 (R&D Systems), or IL-13RA2 (Biolegend) was added and incubated for 2 h on ice with occasional mixing. Cells incubated with isotype antibody served as controls. Cells were washed with PBS/1% BSA prior to the addition of Alexa-Fluor-labeled secondary antibodies. Cells were incubated on ice for an additional 1 h with occasional mixing. After washing with PBS/1% BSA, cells were postfixed with 10% buffered formalin. Cells were analyzed on an Accuri 6 flow cytometer. Data were analyzed using the FCS Express software (DeNovo Software).

Sharma P., Sonawane P., Herpai D., D’Agostino R., Rossmeisl J., Tatter S, & Debinski W. (2020). Multireceptor targeting of glioblastoma. Neuro-oncology Advances, 2(1), vdaa107.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Mouse Eye Tissue

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

Isolated mouse eyes were fixed in 3.7% formaldehyde overnight at 4 °C, cryoprotected in 30% sucrose solution for a minimum of 24 h prior to freezing and 20-μm thick cryosections cut. Sections were incubated in 0.25% Triton X-100 in DPBS buffer (2.7 mM KCl, 1.5 mM KH2PO4, 137.9 mM NaCl, 8.1 mM Na2HPO4–7 H2O [Corning, 21-0310CV]) for 10 min, followed by blocking buffer (5% goat serum, 0.5 g BSA in 50 ml DPBS) for 1 h prior to labeling. Samples were incubated sequentially in primary antibody at 4 °C overnight, followed by fluorescent-conjugated secondary antibody for 1 h at 37°C (Jackson ImmunoResearch Laboratories, 111-295-144, 115-545-003, 115-295-008). Primary antibodies used included: N-cadherin (BD Bioscience, 610921), β-catenin (BD Bioscience, 610154), E-cadherin (Cell Signaling, 24E10), βB-crystallin (Santa Cruz, FL-252), Rac1 (Abcam, ab155938), myosin-II (LifeSpan BioSciences, LS-C84042), WGA (LifeSpan BioSciences, LS-C76576), ZO-1 (Abcam, 61357), EphA2 (R&D, AF639), connexin 50 (ADI, Cx50-A), aquaporin-0 (ADI, AQP01-A), active Rac1 (NewEast Biosciences, 26903), p-myosin (Cell Signaling, 3674), and MLCK (Abcam, ab76092). F-actin was localized with Alexa448-conjugated phalloidin (Invitrogen-Molecular Probes). Nuclei were labeled with TO-PRO-3 (Invitrogen-Molecular Probes).

Logan C.M., Rajakaruna S., Bowen C., Radice G.L., Robinson M.L, & Menko A.S. (2017). N-cadherin regulates signaling mechanisms required for lens fiber cell elongation and lens morphogenesis. Developmental biology, 428(1), 118-134.

+ Open protocol

+ Expand

Immunophenotyping of γδ T Cells

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

The cell phenotype was assessed by fluorescence-activated cell sorting (FACS) by using staining with the monoclonal antibodies MICA, MICB, CD112, CD155, B7-H6 (R&D Systems, clone 875001), and EphA2 (R&D Systems, clone 371805). The tumor cell lines were harvested, washed with cold phosphate-buffered saline (PBS, Sigma) containing 2% FBS, and incubated for 30 min on ice with fluorescently labelled monoclonal antibodies. Gamma-delta T cells were identified in freshly isolated PBMCs labelled with CD3 (Thermo Fisher Scientific, clone SK7), Vδ1 TCR (Thermo Fisher Scientific, clone TS8.2), Vδ2 TCR (BD Pharmingen, clone B6) or Vδ2 TCR (Sony, clone B6). CD27 (BD Pharmingen, clone M-T271), and CD45RA (Exbio, clone MEM-56) were used for immunophenotyping. Samples were washed and acquired using FACSCanto^® (BD Biosciences) and data analyzed using FlowJo^® software (FlowJo, Ashland, OR, USA). Forward and side scatter gating were used to discriminate live cells from dead cells and γδ T cells were derived from SSC vs. FSC-gated bulk PBMCs with doublet exclusion (FSC-A vs. FCS-H). To determine the placement of the gates, appropriate fluorescence minus one (FMO) and unstained controls were used.

Hudecek R., Kohlova B., Siskova I., Piskacek M, & Knight A. (2021). Blocking of EphA2 on Endometrial Tumor Cells Reduces Susceptibility to Vδ1 Gamma-Delta T-Cell-Mediated Killing. Frontiers in Immunology, 12, 752646.

+ 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!