6625-1010 recognises nestin, a large intermediate filament protein originally identified by Hockfield and McKay (1985) It is predominately expressed in stem cells of the developing nervous system. Terminal differentiation is associated with a loss of nestin expression. Nestin expression has also been noted in other embryonic tissues, also in most Glioblastoma multiformes and many melanomas.
Species Cross-Reactivity
Target Species
Cross Reactivity
Human
No
Mouse
Yes
Application
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
Immunofluorescence
Immunohistology - Frozen
1/20 - 1/200
Immunohistology - Paraffin
1/20 - 1/200
Western Blotting
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using the appropriate negative/positive controls.
Immunohistology
We recommend perfusing tissues with 4% paraformaldehyde at pH 7.4 for light microscopy or with either 4% paraformaldehyde at pH 10.0 or 4% paraformaldehyde with 0.1% glutaraldehyde at pH 7.4 for EM. For Immunocytochemistry we recommend using cells fixed in 4% paraformaldehyde buffered with 50 mM sodium borate at pH 9.5.
Western Blotting
6625-1010 reacts with a band at 200-220 kDa in reducing gels of newborn rat or mouse cell extracts. For the purpose of western blotting it is recommended that samples should be boiled in 4 volumes of 125 mM Tris, pH 6.8, 10% 2-mercaptoethanol, 10% glycerol and 4.6% SDS. Membranes should be blocked with milk or BSA. 5% PAGE gels are suggested.
Storage
Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
18 months from date of despatch.
Fusion Partners
Spleen cells from immunised Balb/c mice were fused with cells of the NS1 myeloma cell line
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