MAB345M
Anti-O4 Antibody, clone 81
clone 81 (mAB O4), Chemicon®, from mouse
Synonym(s):
Sulfatide
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
biological source
mouse
Quality Level
100
300
antibody form
purified immunoglobulin
antibody product type
primary antibodies
clone
81 (mAB O4), monoclonal
species reactivity
rat, mouse, human, chicken
manufacturer/tradename
Chemicon®
technique(s)
immunocytochemistry: suitable
immunohistochemistry: suitable
isotype
IgM
suitability
not suitable for Western blot
not suitable for immunoprecipitation
shipped in
wet ice
target post-translational modification
unmodified
Specificity
Recognizes Oligodendrocyte marker O4. Also reacts with certain galactolipids in sperm (see Additional Information library for list).
Application
Anti-O4 Antibody, clone 81 is an antibody against O4 for use in IC, IH with more than 50 product citations.
Immunohistochemistry: 10-20 μg/mL on unfixed, shock frozen tissue.
Immunocytochemistry: 10-20 μg/mL on cells fixed with 4% paraformaldehyde.
Note: O4 is a sulfatide, which can be dissolved out of the membrane by organic solvents; acetone and methanol should not be used for fixation.
Optimal working dilutions must be determined by the end user.
Immunohistochemistry protocol
1. Prepare sections from unfixed, shock frozen tissue. The sections should be 4-5 μm thick. Place the sections on microscope slides.
2. Wash the slide three times for 5 min. each in PBS at room temperature.
3. Block the non-specific binding sites by incubating the sections in a humid chamber with 5% FCS at room temperature for 30 minutes.
4. Wash the slides as described in step 2.
5. Cover the sections with a sufficient amount of MAB345 (10-20 μg/mL in PBS) and incubate in a humid chamber at 37°C for one hour.
6. Wash the slides briefly three times with PBS. Carefully dry around the area to be stained.
7. Cover the sections with a sufficient amount of anti-mouse IgM-fluorescein* solution and incubate in a humid chamber at 37°C for one hour.
8. Wash the slides as described in step 6.
9. Cover the sections with a suitable embedding medium, cover with a cover slip, and examine by fluorescence microscopy.
*HRP or ABC can also be used.
Optimal results can be obtained by titrating the primary and secondary antibodies
Immunocytochemistry
1. Fix the preparations with 4% paraformaldehyde (in PBS) at room temperature for 10 minutes. O4 is a sulfatide which can be dissolved out of the membrane by organic solvents; acetone and methanol should not be used for fixation.
2. Wash the slide three times for 5 min. each in PBS at room temperature.
3. Block the non-specific binding sites by incubating the sections in a human chamber with 5% FCS at room temperature for 30 minutes.
4. Wash the slides as described in step 2.
5. Cover the sections with a sufficient amount of MAB345 (10-20 μg/mL in PBS) and incubate in a humid chamber at 37°C for one hour.
6. Wash the slides briefly three times with PBS. Carefully dry around the area to be stained.
7. Cover the sections with a sufficient amount of anti-mouse IgM-fluorescein solution and incubate in a humid chamber at 37°C for one hour.
8. Wash the slides as described in step 6.
9. Cover the sections with a suitable embedding medium, cover with a cover slip, and examine by fluorescence microscopy.
Note: Do not allow the preparations to dry out during staining.
Immunocytochemistry: 10-20 μg/mL on cells fixed with 4% paraformaldehyde.
Note: O4 is a sulfatide, which can be dissolved out of the membrane by organic solvents; acetone and methanol should not be used for fixation.
Optimal working dilutions must be determined by the end user.
Immunohistochemistry protocol
1. Prepare sections from unfixed, shock frozen tissue. The sections should be 4-5 μm thick. Place the sections on microscope slides.
2. Wash the slide three times for 5 min. each in PBS at room temperature.
3. Block the non-specific binding sites by incubating the sections in a humid chamber with 5% FCS at room temperature for 30 minutes.
4. Wash the slides as described in step 2.
5. Cover the sections with a sufficient amount of MAB345 (10-20 μg/mL in PBS) and incubate in a humid chamber at 37°C for one hour.
6. Wash the slides briefly three times with PBS. Carefully dry around the area to be stained.
7. Cover the sections with a sufficient amount of anti-mouse IgM-fluorescein* solution and incubate in a humid chamber at 37°C for one hour.
8. Wash the slides as described in step 6.
9. Cover the sections with a suitable embedding medium, cover with a cover slip, and examine by fluorescence microscopy.
*HRP or ABC can also be used.
Optimal results can be obtained by titrating the primary and secondary antibodies
Immunocytochemistry
1. Fix the preparations with 4% paraformaldehyde (in PBS) at room temperature for 10 minutes. O4 is a sulfatide which can be dissolved out of the membrane by organic solvents; acetone and methanol should not be used for fixation.
2. Wash the slide three times for 5 min. each in PBS at room temperature.
3. Block the non-specific binding sites by incubating the sections in a human chamber with 5% FCS at room temperature for 30 minutes.
4. Wash the slides as described in step 2.
5. Cover the sections with a sufficient amount of MAB345 (10-20 μg/mL in PBS) and incubate in a humid chamber at 37°C for one hour.
6. Wash the slides briefly three times with PBS. Carefully dry around the area to be stained.
7. Cover the sections with a sufficient amount of anti-mouse IgM-fluorescein solution and incubate in a humid chamber at 37°C for one hour.
8. Wash the slides as described in step 6.
9. Cover the sections with a suitable embedding medium, cover with a cover slip, and examine by fluorescence microscopy.
Note: Do not allow the preparations to dry out during staining.
Physical form
Format: Purified
Purified immunoglobulin in 0.05M Potassium phosphate buffer, pH 8.0 with 0.3M NaCl and 0.05% sodium azide.
Analysis Note
Control
Rat cortical stem cells or day 3 cell cultures of brains from mouse embryos
Rat cortical stem cells or day 3 cell cultures of brains from mouse embryos
Other Notes
Concentration: Varies, see lot specific CoA
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
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Systemic injection of neural stem/progenitor cells in mice with chronic EAE.
Donega, M; Giusto, E; Cossetti, C; Schaeffer, J; Pluchino, S
Journal of Visualized Experiments null
Marc-André Mouthon et al.
Journal of neurochemistry, 99(3), 807-817 (2006-08-24)
Developing and adult forebrains contain neural stem cells (NSCs) but no marker is available to highly purify them. When analysed by flow cytometry, stem cells from various tissues are enriched in a 'side population' (SP) characterized by the exclusion of
Yifat Amir-Levy et al.
Multiple sclerosis international, 2014, 926134-926134 (2015-01-23)
Background. The neural stem cells (NSCs) migrate to the damaged sites in multiple sclerosis (MS) and in experimental autoimmune encephalomyelitis (EAE). However, the differentiation into neurons or oligodendrocytes is blocked. Epidermal growth factor (EGF) stimulates NSC proliferation and mobilization to
Differentiation of human breast-milk stem cells to neural stem cells and neurons.
Hosseini, SM; Talaei-Khozani, T; Sani, M; Owrangi, B
Neurology research international null
Zhuo Wang et al.
Development, growth & differentiation, 53(3), 357-365 (2011-04-12)
We attempted to test whether the differentiated NIH/3T3 fibroblasts could be differentiated into neuronal cells without any epigenetic modification. First, a neurosphere assay was carried out, and we successfully generated neurosphere-like cells by floating cultures of NIH/3T3 fibroblasts in neural
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