In neonatal rat glia MAB328 stains oligodendrocytes and does not react with type 1 astrocytes, type 2 astrocytes or fibroblasts. Does not stain cultures of rat Schwann cells. In tissue sections MAB328 labels oligodendrocytes and CNS myelin with no labeling of peripheral myelin. MAB328 detects MOSP which is expressed at day 4-5 of neonatal rat, which is about 1-2 after the appearance of GC or sulfatide. The initial expression of MOSP occurs at the stage in development when oligodendrocytes are elaborating processes and just beginning to form membrane sheets (Mu QQ & C Dyer, 1994).

Rat glial membranes and whole brain white matter.

Immunocytochemistry

Immunohistochemistry at 1:1,000-1:5000

Not suggested for use in Western blot.

Optimal working dilutions must be determined by end user.

IMMUNOHISTOCHEMISTRY PROTOCOL FOR MAB328

This antibody has been used successfully on 30 mm, free floating, 4% paraformaldehyde fixed rat brain tissue. All steps are performed under constant agitation. Suggested protocol follows.

1) 3 x 10 minute washes in TBS (with or without 0.25% Triton).

2) Incubate for 30 minutes in TBS with 3% serum (same as host from secondary antibody).

3) Incubate primary antibody diluted appropriately in TBS with 1% serum (same as host from secondary antibody) (with or without 0.25% Triton) for 2 hours at room temperature followed by 16 hours at 4°C.

4) 3 x 10 minute washes in TBS.

5) Incubate with secondary antibody diluted appropriately in TBS with 1% serum (same as host from secondary antibody).

6) 3 x 10 minute washes in TBS.

7) ABC Elite (1:200 Vector Labs) in TBS.

8) 2 x 10 minute washes in TBS.

9) 1 x 10 minute wash in phosphate buffer (no saline).

10) DAB reaction with 0.06% NiCl added for intensification.

11) 2 x 10 minute washes in PBS.

12) 1 x 10 minute wash in phosphate buffer (no saline).

Research Category
Neuroscience

Research Sub Category
Neuronal & Glial Markers

This Anti-Oligodendrocytes Antibody, clone CE-1 is validated for use in IP, IC, IH, IH(P) for the detection of Oligodendrocytes.

Maintain at -20°C in undiluted aliquots for up to 12 months. Avoid repeated freeze/thaw cycles.

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.