Antibodies for CNS-resident cell staining were from the following companies: anti-GFAP (2A5) from StemCell Technologies (BC, Canada); anti-NeuN from Millipore (Billerica, MA); anti-CD11b, antiA2B5 and anti-OSP (oligodendrocyte-specific protein) from Abcam (Cambridge, MA) and antibody for IL-9R from Biolegend (San Diego, CA). Fluorescent-conjugated secondary antibodies were from Jackson ImmunoResearch Laboratories (West Grove, PA). Recombinant IL-9, IL-17, IL-1β, IFN-γ, TNF-α, PDGF, bFGF, NT3 and M-CSF were from PeproTech (Rocky Hill, NJ). T3 and T4 were from Sigma-Aldrich (St. Louis, MO).
Anti a2b5
Manufactured by Abcam
Sourced in Morocco, United States
Anti-A2B5 is a primary antibody that recognizes the A2B5 antigen, a cell surface marker expressed on glial precursor cells and subsets of neurons. It is commonly used in research applications to identify and isolate these cell types.
Automatically generated - may contain errors
Lab products found in correlation
Anti gfap, by Merck Group (2 mentions)
Fluorescent conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions)
Il 1β, by Thermo Fisher Scientific (1 mentions)
M csf, by Thermo Fisher Scientific (1 mentions)
Il 17, by Thermo Fisher Scientific (1 mentions)
Anti neun, by Merck Group (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Recombinant il 9, by Thermo Fisher Scientific (1 mentions)
Anti cd11b, by Abcam (1 mentions)
Anti brdu, by Merck Group (1 mentions)
Anti nestin, by Merck Group (1 mentions)
Anti s1p1, by R&D Systems (1 mentions)
Anti β tubulin 3, by Merck Group (1 mentions)
Anti olig2, by Abcam (1 mentions)
Anti ki67, by Cell Signaling Technology (1 mentions)
Anti sox2, by Merck Group (1 mentions)
Anti olig2, by Merck Group (1 mentions)
Anti mbp, by Bio-Rad (1 mentions)
3 protocols using anti a2b5
1
Immunostaining of CNS Cell Types
2
Immunostaining of Neural Stem Cells
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After 30 min fixation in 4% paraformaldehyde, cells were washed three times with 0.01 M PBS and incubated in 0.5% Triton-100. 1% BSA was used prior to an overnight incubation with the primary antibodies. Anti-Brd-U (1 : 800, Sigma, USA), anti-Ki67 (1 : 200, Cell Signaling), and anti-Nestin (1 : 400, Millipore, USA) were used as stem cell markers. Anti-β-tubulin III (1 : 800, Sigma, USA), anti-GFAP (1 : 160, Sigma, USA), anti-A2B5 (1 : 200, Abcam, USA), anti-Olig2 (1 : 500, Abcam, USA), and anti-CC1 (1 : 200, Life Science Research, USA) were used as specific cell type makers. Anti-S1P1 (1 : 100, R&D System, USA) or anti-S1P2 (1 : 200, Novus Biologicals, USA) were used to confirm the expression of S1P receptors in NSCs. FITC-conjugated goat secondary antibodies (1 : 100, Zhongshan, China) and TRITC-conjugated goat secondary antibodies (1 : 100, Zhongshan, China) were used for labeling the different source of primary antibodies separately. Subsequently, the samples were stained with DAPI nuclei staining prior to confocal laser scanning microscopy (Leica, SP2, Germany).
3
Immunocytochemistry of Neural Cell Markers
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Cells were washed with PBS and fixed with 4% paraformaldehyde (PFA) for 10 min at room temperature. After fixation, cells were washed three times with PBS. Cells were then blocked at least for 1 h in blocking solution (1% BSA, 3% goat serum, 0.1% Triton in PBS, unless stated otherwise). The blocking solution was removed and the primary antibody (anti-OLIG2 1:400, EMD Millipore, catalog no. 2367; anti-O4 1:200, Immunosolv [product discontinued]; anti-MBP 1:250, Bio-Rad, catalog no. MCA409S; anti-SOX2 1:100, Sigma, catalog no. S9072; anti-SOX9 1:100, EMD Milipore, catalog no. AB5535; anti-GFAP 1:1,000, Sigma, catalog no. G9269; anti-NESTIN 1:10, DSHB Hybridoma, catalog no. rat-401; anti-TUJ1 1:500, BioLegend, catalog no. 801202; anti-A2B5 1:100, Abcam, catalog no. ab53521) was added and incubated at 4°C overnight in blocking solution. After staining with primary antibody, cells were washed three times with PBST for 15 min. Cells were then stained with an appropriate secondary antibody (Alexa Fluor range; 1:1,000 dilutions) for at least 1 h in the dark at room temperature. Cells were washed two times with PBS, stained with DAPI (1:5,000 in PBS) for at least 5 min and washed with PBS twice.
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