RE: Oct-2 Antibody?Oct-2 (Ab-1) Cat# NA44 Oncogene Research Prod ucts

From:"Ward, Elizabeth"

This may help....

Oct-2 (Ab-1) Cat# NA44
BACKGROUND
The Oct proteins make up a family of transcription
factors that contain bipartite DNA-binding POU
domains, which in turn are members of the helix-turnhelix
family of DNA binding domains (1,2). Oct-1 and
Oct-2 are transcription factors that bind to the same
octamer motif, ATGCAAAT, but possess different
transcriptional activities and expression patterns (3,4).
The Oct elements are the most conspicuous regulatory
element in B cells and are found in every Ig promoter
and in most Ig enhancers (5). While the Oct-1 protein is
found ubiquitously expressed, Oct-2 (also known as
OTF2, NF-A2, and Oct-2b) appears to be restricted in
expression to B-cells, neuronal cells, and some T cells
(3,4, and Reviewed in 6). At least in B-cells, the Oct
proteins bind to DNA together with another protein
transcription factor, octamer binding factor-1 (OBF-1),
to form a ternary complex (2,7). This latter observation
suggests that DNA binding factors exhibiting cell
restricted expression may also have co-factors thereby
adding to their specificity in activity. Oct-2 undergoes
alternative splicing and multiple protein isoforms are
expressed in both B cells and neuronal cells (4).
Alternative splicing of Oct-2 generates proteins with
altered promoter selectivity (8). Neuronal cells
predominantly express two isoforms of Oct-2 that
appear to act as transcriptional repressors whereas the
isoforms predominantly seen in B cells activate
transcription (9). This dual capability of Oct-2 appears
to derive from the presence of distinct activation and
repression domains within the protein along with the
fact that Oct-2 functions as a multimer with individual
multimers composed of one or more isoforms
(Reviewed in 6). Oct-2 expression in neurons can be
induced by NGF, and the isoforms induced have
transcriptional repression activities; thus, neuronal Oct-
2 appears to be an immediate early gene whose
expression leads to transcriptional repression (10).
ORIGIN
Clone PT1 is a mouse monoclonal antibody generated
by immunizing mice with recombinant Oct-2 (11) and
fusing the splenocytes with Sp2/0 mouse myeloma cells.
CHARACTERISTICS
,VRW\SHIgG1
(SLWRSHNH-terminal segment of Oct-2 present in all
isoforms (4, 11).
KNOWN SPECIES REACTIVITY
3RVLWLYHHuman
Negative: Mouse
FORMULATION
100 mg of purified antibody in 1.0 mL of 0.05 M
sodium phosphate buffer containing 50 % glycerol.
STORAGE
Store at -20C. For long term storage, aliquot into
smaller volumes and store at -20oC. Avoid multiple
freeze/thaw cycles. If stored under proper conditions,
product guaranteed until expiration date stated.
COMMENTS
NA44 can be used in electrophoretic mobility shift
assays as described (11).
APPLICATIONS
Amt Label Pretreatment Detection Positive
Control
Negative
Control
Immunoprecipitation
1 mg 35S or
IP/blot (11)
Metabolic
labeling
Autoradiography or
blotting
Daudi cells HeLa cells
Western Blot* 1 g/mL NA None Chemiluminescence Daudi cells HeLa cells
Frozen Sections NT
Paraffin Sections NT
Immunofluorescence
NT
#See NOTES NA: Not applicable NT: Not tested
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ONCOGENE RESEARCH PRODUCTS ORDERING INFORMATION
650 Albany Street Research reagents may be ordered by mail at
Boston, MA 02118 the address shown at left, by telephone,
Tel: 617 536-2223 800 662-2616 E-mail, or fax or from your local
Fax: 617 536-9911 800 828-4871 Oncogene Research Products supplier.
E-Mail: oncogene@apoptosis.com Terms are net 30 days F.O.B.
Web Page: http://www.apoptosis.com shipping point.
http://www.neuroproducts.com
For research use only, not for use in diagnostic procedures
Oct-2 (Ab-1) Cat# NA44
*(1(5$/5()(5(1&(6
1. Herr, W., et al. (1988) Genes and Dev. 2: 1513-
1516
2. Aurora, R., and Herr, W. (1992) Mol. Cell. Biol.
12: 455-467
3. Sturm, R.A., et al. (1988) Genes Dev. 2: 1582-
1599
4. Wirth, T., et al. (1991) Nuc. Acids Res. 19: 43-51
5. Staudt, L.M., and Lenardo, M.J. (1991) Ann. Rev.
Immunol. 9: 373-396
6. Latchman, D.S. (1996) Int. J. Biochem. Cell Biol.
28: 1081-1083.
7. Strublin, M., et al. (1995) Cell 80: 497-506
8. Tanaka, M., et al. (1992) Cell 68: 755-767.
9. Lillycrop, K.A. and Latchman, D.S. (1992) J.
Biol. Chem. 267: 24960-24965.
10. Kendall, G., et al. (1995) J. Neurosci Res. 40:
169-176.
11. Lai, J.-S. & Herr, W. (1992) Proc. Natl. Acad. Sci.
USA 89: 6958-6962.
12. Scheidereit, C., et al. (1987). Cell 51: 783-793.
13. Clerc, R.G., et al. (1988) Genes Dev. 2: 1570-
1581.
NOTES: Oct-2 can be detected in 50 - 100 mg of Daudi cell lysate by western
blotting with NA44. Other
cell lines known to express Oct-2 include WEHI231, BJA-B, HAFTL 1, and
X63Ag8 (4,12,13). HeLa and
293 cells (11) do not express detectable levels of Oct-2 protein. The
predominant band detected is 55 - 60
kDa which may appear as a doublet due to expression of additional isoforms.
A second band of approx. 75
kDa, corresponding to the Oct2-5/Oct-2b isoform (4,11), is also detected,
albeit at reduced amounts with
NA44 but not with NA45. Careful titration of the secondary detector antibody
may be necessary for
optimum signal.

-----Original Message-----
From: Drew Sally A. [mailto:sa.drew@hosp.wisc.edu]
Sent: Thursday, February 21, 2002 1:16 PM
To: 'Histonet'
Subject: Oct-2 Antibody?


Are any of you wonderful people familiar with an antibody called Oct-2?  We
have a hematopathologist asking us what it is...




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