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)).
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.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 700 confocal microscope, by Zeiss (2 mentions)
Appropriate fluorescent secondary antibodies, by Jackson ImmunoResearch (1 mentions)
Dapi vectorshield mounting medium, by Vector Laboratories (1 mentions)
Alexa fluor 488 donkey anti goat, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Mouse anti cxcr4, by R&D Systems (1 mentions)
Rabbit anti β catenin, by Cell Signaling Technology (1 mentions)
Rabbit anti cdx2, by Abcam (1 mentions)
Mouse anti cd44, by BD (1 mentions)
Rabbit anti cd133, by Abcam (1 mentions)
2 protocols using goat anti epha2
1
Immunolocalization of EphA2 and ephrin-A5 in mouse lens
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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