Recombinant Human
STAT1
PRODUCT ANALYSIS SHEET
Catalog Number:
PHF0011
Lot Number:
1397177
Quantity:
100 g
Molecular Weight:
~ 90 kDa
Purity:
>95% as determined by SDS-PAGE analysis.
Biological Activity:
Recombinant human STAT1 is phosphorylatable in vitro, using either
immunoprecipitated JAK1 or JAK2, or whole cell extracts derived from interferon-γstimulated HEK 293 cells. This phosphorylation can be monitored by Western blot
analysis using BioSource’s phosphorylation site specific antibody directed to STAT
[pY701] (Catalog # 44-378), in conjunction with chemiluminescence detection
methods. Optimization of the cell stimulation protocol, cell lysis procedure, and
reaction conditions may be required for each specific application.
Formulation:
Lyophilized, carrier protein-free.
Sterility:
Filtered through a 0.22 micron sterile filter.
Production:
Recombinant Human STAT1 was produced in E. coli with an amino-terminal
polyhistidine tag and purified via sequential chromatography.
Reconstitution
Recommendation:
We recommend that this vial be briefly centrifuged prior to opening to bring the
contents to the bottom. Reconstitute lyophilized recombinant human STAT1 in
10 mM Tris, pH 7.5, to a concentration of 0.2-1.0 mg/mL.
Suggested Working
Dilutions:
The optimal concentration should be determined for each specific application.
Expiration Date:
Expires one year form date of receipt when stored as instructed.
Storage:
Store lyophilized recombinant human STAT1 at 2-8C, preferably desiccated. Upon
reconstitution, apportion the solution into working aliquots. STAT1 solutions are
stable at 4C for up to 2 weeks. For maximal stability, store the aliquots at <-20C.
Avoid repeated freeze/thaw cycles.
References:
Schindler, C., X.Y. Fu, T. Improta, R. Aebersold, and J.E. Darnell Jr. (1992) Proteins
of transcription factor ISGF-3: One gene encodes the 91- and 84-kDa ISGF-3
proteins that are activated by interferon alpha. Proc. Nat’l. Acad. Sci. U.S.A.
89:7836-7839.
Quelle, F.W., W. Thierfelder, B.A. Witthuhn, B. Tang, S. Cohen, and J.N. Ihle (1995)
Phosphorylation and activation of the DNA binding activity of purified STAT1 by the
This product is for research use only. Not for use in diagnostic procedures.
www.invitrogen.com
Manufactured under ISO 13485 Quality Standard
Invitrogen Corporation • 542 Flynn Rd • Camarillo • CA 93012 • Tel: 800.955.6288 • E-mail: techsupport@invitrogen.com
PI PHF0011
(Rev 1.1) (DCC-08-1232)
Important Licensing Information - These products may be covered by one or more Limited Use Label Licenses (see the Invitrogen Catalog or our website,
www.invitrogen.com). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated,
these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.
Janus protein-tyrosine kinases and the epidermal growth factor receptor. J. Biol.
Chem. 270:20775-20780.
References (cont’d):
Greenlund, A.C., M.O Morales, B.L. Viviano, H. Yan, J. Krolewski, and R.D.
Schrieiber (1995) STAT recruitment by tyrosine-phosphorylated cytokine receptors:
An order reversible affinity-driven process. Immunity 2:677-687.
Vignais, M-L. and M. Gilman (1999) Distinct mechanisms of activation of STAT1
and STAT3 by platelet-derived growth factor receptor in a cell-free system. Mol. Cell
Biol. 19:3727-3735.
Wang, Y., T.R. Wu, S. Cai, T. Welte, and Y.E. Chen (2000) STAT1 as a component
of tumor necrosis factor alpha receptor 1-TRADD signaling complex to inhibit
NF-kappaB activation. Mol. Cell. Biol. 20:4505-4512.
Bromberg, J. and X. Chen (2001) STAT proteins: Signal transducers and activators of
transcription. Methods Enzymol. 333:138-151.
Nair, J.S., C.J. DaFonseca, A. Tjernberg, W. Sun, J.E. Darnell Jr., B.T. Chait, and J.J.
Zhang (2002) Requirement of Ca2+ and CaMKII for STAT1 Ser-727 phosphorylation
in response to IFN-gamma. Proc. Nat’l. Acad. Sci. U.S.A. 99:5971-5976.
Levy, D.E. and J.E. Darnell Jr. (2002) STATs: Transcriptional control and biological
impact. Nat. Rev. Mol. Cell. Biol. 3:651-662.
kDa
100-
1
2
75-
1
2
- STAT1
50STAT1 pan
antibody
STAT1 [pY701]
antibody
(Cat. # 44-360)
(Cat. # 44-378)
In vitro activation of recombinant STAT1. Western blot analysis of recombinant STAT1 incubated with
unstimulated (Lane 1) and IFN--stimulated (Lane 2) HEK 293 cell extracts in a standard kinase reaction.
Recombinant STAT1 was purified by batch binding to Ni-NTA agarose, washed, and eluted with SDS-sample
buffer and electrophoresed on a 7.5% SDS PAGE gel. Proteins were transferred to PVDF membranes and
incubated with 1 g/mL of STAT1 pan antibody (Catalog # 44-360) or STAT1 [pY701] phosphorylation site
specific antibody (Catalog # 44-378). The signal was detected using a goat anti-rabbit IgG alkaline phosphatase
conjugated antibody (Catalog # ALI4405) at a 1:5000 dilution and the membrane was incubated with
CDP-substrate using the WesternStarTM method (Tropix). The membrane was then exposed to Kodak BioMax
film.
For detailed procedures and reaction conditions, please see:
http://www.biosource.com/content/catalogContent/tds3/moreinfo/STAT1%20activation1.pdf
This product is for research use only. Not for use in diagnostic procedures.
www.invitrogen.com
Manufactured under ISO 13485 Quality Standard
Invitrogen Corporation • 542 Flynn Rd • Camarillo • CA 93012 • Tel: 800.955.6288 • E-mail: techsupport@invitrogen.com
PI PHF0011
(Rev 1.1) (DCC-08-1232)
Important Licensing Information - These products may be covered by one or more Limited Use Label Licenses (see the Invitrogen Catalog or our website,
www.invitrogen.com). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated,
these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.