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Goat anti epha2

Manufactured by R&D Systems
Sourced in United States

Goat anti-EphA2 is a polyclonal antibody produced in goats and specifically targets the EphA2 receptor. EphA2 is a member of the Eph receptor tyrosine kinase family and plays a role in various cellular processes. This antibody can be used for the detection and analysis of EphA2 in research applications.

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2 protocols using goat anti epha2

1

Immunolocalization of EphA2 and ephrin-A5 in mouse lens

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Lens capsule flat mounts with attached epithelial and peripheral fiber cells were prepared from postnatal day 21 (P21) non-GFP WT mouse lenses as previously described (Cheng et al., 2013 (link); Cheng and Gong, 2011 (link)). Briefly, freshly dissected lenses were flash fixed in cold 100% methanol for 45 seconds. Lens capsules (along with attached epithelial and peripheral fiber cells) was gently dissected away from the fiber cell mass using radial cuts. Capsules were blocked (10% normal donkey serum and 0.3% Triton X-100) for 1 hour at room temperature before staining with goat anti-EphA2 (R&D Systems) and rabbit anti-ephrin-A5 (Invitrogen) primary antibody overnight at 4°C. Primary antibody incubation was followed washing and then incubation in appropriate fluorescent secondary antibodies (Jackson ImmunoResearch Laboratories) for 2 hours at room temperature. Lens capsules were then washed and flat mounted with DAPI VectorShield mounting medium (Vector Laboratories, Inc.). Z-stack images were collected by a Zeiss LSM700 confocal microscope, and 3D images were reconstructed from z-stack data collected at 0.38 μm steps using the ZEN software. Antibody specificity was tested previously using knockout tissues (Cheng and Gong, 2011 (link)).
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2

Immunofluorescence staining of cancer stem cell markers

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For immunofluorescence staining on tissue sections, CCSCs, mCCSCs and cCCSCs cells antibodies/antisera used were: mouse anti-Human Nuclei (1:100, Millipore-Merk KGaA, Darmstadt, Germany), mouse anti-BMI1 (1:100, Merk Millipore), rabbit anti-EpCAM (1:50, Cell Signaling, Beverly, MA, USA), rabbit anti-ALDHA1 (1:50, Cell Signaling), rabbit anti-bCatenin (1:50, Cell Signaling), rabbit anti-CD133 (1:100, AbCam, Cambridge, UK), rabbit anti-OLFM4 (1:200, AbCam), rabbit anti-CDX2 (1:100, AbCam), rabbit anti-CK20 (1:100, Abcam), rabbit anti-Lgr5 (1:50, Merk SigmaAldrich), rabbit anti-Laminin (1:400, Merk SigmaAldrich), mouse anti-Vimentin (1:100, Agilent, Santa Clara, CA, USA), mouse anti-HLA-ABC (1:100, Agilent), goat anti-EphA2 (1:50, R&D System, Minneapolis, MN, USA), mouse anti-CXCR4 (1:100, R&D System), rabbit anti-KI67 (1:200, Leica Microsystem, Wetzlar, Germany), mouse anti-CD44 (1:50, BD Biosciences, San Jose, CA, USA), rabbit anti-Wnt5a (1:50, LS Biosciences, Seattle WA, USA), mouse anti-MUC2 (1:50, Novusbio, Littleton, CO, USA). Goat anti mouse AlexaFluor488/546 (1:1000, Thermo Fisher Scientific, Waltham, MA, USA), goat anti rabbit AlexaFluor488 (1:1000, Thermo Fisher Scientific) and donkey anti goat AlexaFluor488 (1:1000, Thermo Fisher Scientific) secondary antibodies were used to visualize the primary antibody staining.
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