We killed eight rats per group from the six groups (sham+ vehicle, sham +BMMNC, 2VO +vehicle, 2VO +BMMNC, 2VO+VEGF+BMMNC, 2VO+SU5416+BMMNC) to assess BMMNC migration on day 1 after BMMNC transplantation. The brains and left lower lungs of each rat were cut into 25 μm sections and incubated with anti-eGFP antibody (1:1000, Abcam, Cambridge, MA, USA) via the methods described above. All sections were mounted on coverslips with a drop of mounting medium containing 1.5 μg/mL 4′,6-diamidino-2-phenylindole (DAPI; Santa Cruz Biotechnology).
Anti egfp antibody
Manufactured by Abcam
Sourced in United States, United Kingdom
Anti-EGFP antibody is a laboratory reagent used to detect and quantify the presence of Enhanced Green Fluorescent Protein (EGFP) in samples. It provides a specific and sensitive tool for researchers working with EGFP-tagged proteins or cells.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Santa Cruz Biotechnology (1 mentions)
Veletta ccd digital camera, by Thermo Fisher Scientific (1 mentions)
Bond polymer refine detection kit, by Leica (1 mentions)
Goat on rodent hrp polymer, by Biocare Medical (1 mentions)
Bond rx, by Leica (1 mentions)
Dako protein block serum free, by Agilent Technologies (1 mentions)
4 protocols using anti egfp antibody
1
Tracking BMMNC Migration in Rat Brain
2
EGFP Immunoprecipitation and Western Blotting
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Cell lysates contain at least 1000 μg of protein from each sample were pre-cleared with 50 μl of protein A-Sepharose beads for 1 h at 4°C and incubated with 5 μg of anti-EGFP antibody (Abcam, Cambridge, UK) overnight at 4°C on a rotator. The total volume of this reaction was 1ml. Following antibody incubation, 100 μl of protein A sepharose beads (50% slurry) were added and rotated at 4°C for 3 h. The beads were then centrifuged at 12,000g for 5min and washed for 5 times with 1% NP40 lysis buffer. The precipitates were eluted by adding of 50 μl of 1× SDS-PAGE sample loading buffer (50 mm Tris-HCl, pH 6.8, 100 mm DTT, 2% SDS, 0.1% bromphenol blue, 10% glycerol), followed by boiling at 100°C for 5 min. The supernatant obtained after centrifugation was resolved by 10% SDS-PAGE and subjected to Western blot analysis.
3
Ultrastructural Localization of UPIb in MDCK and HeLa Cells
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The MDCK and HeLa cells transfected with UPIb-EGFP or UPIb/UPIIIa were fixed as described previously66 (link),67 . Cells were incubated with the anti-EGFP antibody (1:250; Abcam), washed in PBS and incubated in anti-rabbit Fab’ fragments coupled to 1.4-nm nanogold (1:100; Molecular Probes). Nanogold was enlarged by the GoldEnhance protocol (Nanoprobes Inc., Yaphank, NY, USA). Sections of 65-nm thickness were collected and EM images were acquired by a TechaiTM-G2 Spirit electron microscope equipped with a VELETTA CCD digital camera (FEI, Eindhoven, The Netherlands).
4
Visualizing EGFP Expression via IHC
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IHC, conducted to visual EGFP protein expression in injection site and LN tissues, was performed on formalin-fixed paraffin embedded sections using the Leica Bond RX autostainer (Leica Microsystems, Buffalo Grove, IL). Sections were baked and deparaffinized on the instrument, followed by an epitope retrieval for 10 min using Leica Epitope Retrieval Buffer 1 (catalog no. AR9961). Dako serum-free protein block (catalog no. X090930-2, Agilent Dako, Santa Clara, CA) was incubated on the slides for 15 min at room temperature. Anti-EGFP antibody (catalog no. ab6673, or ab290 Abcam, Cambridge, UK) was used at 2.4 or 0.3 μg/mL dilution at room temperature for 30 min. Secondary antibody and detection were performed using the Goat-on-Rodent HRP-Polymer (catalog no. GHP516, BioCare Medical, Pacheco, CA) and/or the Bond Polymer Refine Detection Kit (catalog no. DS9800, Leica Microsystems). Bond DAB Enhancer (catalog no. AR9432, Leica Microsystems) and bluing reagent (catalog no. 3802918, Leica Microsystems) were used to enhance the color.
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