CTB488 was injected into the duodenum, ileum, and proximal colon. Mice were sacrificed by cervical dislocation and the CG-SMG and NG were harvested and fixed overnight in 4% PFA. Tissue was then washed four times in DPBS at RT and permeabilized in PTxwH for 4 hr at RT. Primary antibody anti-AlexaFluor488 (1:400, Thermo Fisher Scientific, A-11094) was added to the samples in PTxwH and incubated at 4°C for 48 h. Samples were washed four times in PTxwH at RT and then stained with goat-anti rabbit AF555/568/647 at 4°C for 24 h. Samples were washed four times in PTxwH at RT, covered in Fluormount G, and coverslipped for confocal imaging. Each ganglion was captured in full by multiple z-stacks and the total number of CTB+ neurons were counted.
Anti alexafluor488
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Anti-AlexaFluor488 is a secondary antibody used in immunofluorescence and other immunoassay applications. It is designed to specifically bind and detect the AlexaFluor488 fluorescent label.
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Lab products found in correlation
4 protocols using anti alexafluor488
1
Retrograde Tracing of Intestinal Neurons
2
Antibodies for EGFR Endocytosis Analysis
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rabbit polyclonal anti-EGFR (epitope: aa 1172-1186, Homo sapiens), mouse anti-eps15 (epitope: aa 2-330, Mus musculus), rabbit anti-eps15L1 (epitope: aa 216-266, Mus musculus), mouse anti-epsin1/2 (epitope: aa 249-401 of epsin1, Homo sapiens) were produced in-house and used in IB experiments. Mouse anti-EGFR 13A9 (Genentech) was used to follow endocytosis in in vivo EM analyses. Anti-Alexa Fluor 488 (ThermoFisher) was used for cryosection immunolabeling for EM analysis. Other antibodies used in IB are listed in the Key Resources Table.
3
Characterization of hDPSCs by Flow Cytometry
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The characterization of the hDPSCs using flow cytometry was based on the expression analysis of cell surface molecules CD14 (Anti-CD14 FITC Alexa Fluor 488 - Thermo Fisher Scientific, Waltham, Massachusetts, USA), CD45 (Anti-Alexa Fluor 488 - Thermo Fisher Scientific, Waltham, Massachusetts, USA) CD105 (anti-CD105 Alexa Fluor 488 - Thermo Fisher Scientific, Waltham, Massachusetts, USA), yielding 30,000 events per tested sample.
Sixth-passage cells were used, thawed, cultured and trypsinized to reach 80% to 90% confluence in the 25-cm2 flask. After centrifugation at 462 g for 10 minutes, the cells were counted in a Neubauer chamber and centrifuged again at 462 g for 10 minutes. The pellet was re-suspended in 250 µL of PBS divided into five aliquots with 50 µL each, in 15 ml Falcon tubes. The four antibodies were each placed in a tube (CD14 – 10 µL, CD45 – 10 µL, CD105 – 5 µL). One tube was used as the control. The tubes were left in an oven at 37°C with 5% CO2 and 95% humidity for 30 minutes without light. After this period, two centrifugations at 462 g were performed for 5 minutes. 400 µL of PBS was added to the resulting pellet. After this, they were transferred to a cytometer tube, where data acquisition was conducted followed by analysis in a flow cytometer. The results were plotted in a histogram for evaluation.
Sixth-passage cells were used, thawed, cultured and trypsinized to reach 80% to 90% confluence in the 25-cm2 flask. After centrifugation at 462 g for 10 minutes, the cells were counted in a Neubauer chamber and centrifuged again at 462 g for 10 minutes. The pellet was re-suspended in 250 µL of PBS divided into five aliquots with 50 µL each, in 15 ml Falcon tubes. The four antibodies were each placed in a tube (CD14 – 10 µL, CD45 – 10 µL, CD105 – 5 µL). One tube was used as the control. The tubes were left in an oven at 37°C with 5% CO2 and 95% humidity for 30 minutes without light. After this period, two centrifugations at 462 g were performed for 5 minutes. 400 µL of PBS was added to the resulting pellet. After this, they were transferred to a cytometer tube, where data acquisition was conducted followed by analysis in a flow cytometer. The results were plotted in a histogram for evaluation.
4
Retrograde Tracing of Intestinal Neurons
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