Revision 6

#14995Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-F, IF-IC, ChIP, ChIP-seq

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

84, 91

Source/Isotype:

Rabbit IgG

UniProt ID:

#P42224

Entrez-Gene Id:

6772

Product Information

Product Usage Information

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:400 - 1:1600
Immunofluorescence (Frozen) 1:400 - 1:1600
Immunofluorescence (Immunocytochemistry) 1:400 - 1:1600
Chromatin IP 1:50
Chromatin IP-seq 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #84689.

Specificity / Sensitivity

Stat1 (D4Y6Z) Rabbit mAb recognizes endogenous levels of total Stat1 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to a region near the carboxy terminus of human Stat1 protein.

Background

The Stat1 transcription factor is activated in response to a large number of ligands (1) and is essential for responsiveness to IFN-α and IFN-γ (2,3). Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization, nuclear translocation, and DNA binding (4). Stat1 protein exists as a pair of isoforms, Stat1α (91 kDa) and the splice variant Stat1β (84 kDa). In most cells, both isoforms are activated by IFN-α, but only Stat1α is activated by IFN-γ. The inappropriate activation of Stat1 occurs in many tumors (5). In addition to tyrosine phosphorylation, Stat1 is also phosphorylated at Ser727 through a p38 mitogen-activated protein kinase (MAPK)-dependent pathway in response to IFN-α and other cellular stresses (6). Serine phosphorylation may be required for the maximal induction of Stat1-mediated gene activation.

  1. Heim, M.H. (1999) J Recept Signal Transduct Res 19, 75-120.
  2. Durbin, J.E. et al. (1996) Cell 84, 443-50.
  3. Meraz, M.A. et al. (1996) Cell 84, 431-42.
  4. Ihle, J.N. et al. (1994) Trends Biochem Sci 19, 222-7.
  5. Frank, D.A. (1999) Mol Med 5, 432-56.
  6. Wen, Z. et al. (1995) Cell 82, 241-50.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalStain is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 6
#14995

Stat1 (D4Y6Z) Rabbit mAb

Western Blotting Image 1: Stat1 (D4Y6Z) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using Stat1 (D4Y6Z) Rabbit mAb.
Western Blotting Image 2: Stat1 (D4Y6Z) Rabbit mAb Expand Image
Western blot analysis of extracts from A549 cells (lane 1) or STAT1 knock-out cells (lane 2) using Stat1 (D4Y6Z) Rabbit mAb #14995 (upper), and β-actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the STAT1 knock-out A549 cells confirms specificity of the antibody for STAT1.
Western Blotting Image 3: Stat1 (D4Y6Z) Rabbit mAb Expand Image
Western blot analysis of extracts from whole lung from wild type 129S6/SvEvTac mice (model #129SVE-F) (lane 1), or Stat1 knock-out mice (model #2045-F) (lane 2), whole spleen extracts from wild type 129S6/SvEvTac mice (lane 3), or Stat1 knock-out mice (lane 4), whole liver extracts from wild type 129S6/SvEvTac mice (lane 5), or Stat1 knock-out mice (lane 6) using Stat1 (D4Y6Z) Rabbit mAb #14995 (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). The truncated protein observed in the knock-out tissues is the result of a targeted homozygous mutation in the Stat1 gene (4), and confirms the specificity of the antibody for Stat1. Mice from Taconic Biosciences, Inc.
Immunoprecipitation Image 1: Stat1 (D4Y6Z) Rabbit mAb Expand Image
Immunoprecipitation of Stat1 from MCF7 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Stat1 (D4Y6Z) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using Stat1 (9H2) Mouse mAb #9176.
Immunohistochemistry Image 1: Stat1 (D4Y6Z) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Stat1 (D4Y6Z) Rabbit mAb.
Immunohistochemistry Image 2: Stat1 (D4Y6Z) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded lung (left), spleen (middle), and liver (right) from wild type 129S6/SvEvTac mouse (model #129SVE-F) (top) or Stat1 knock-out mouse (model #2045-F) (bottom) using Stat1 (D4Y6Z) Rabbit mAb. The targeted mutation in the KO animal results in a reduction of expression and truncation of the protein(4). Mice from Taconic Biosciences, Inc.
Immunofluorescence Image 1: Stat1 (D4Y6Z) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed, frozen thymus from wild type 129S6/SvEvTac mouse (model #129SVE-F) (left) or Stat1 knock-out mouse (model #2045-F) (right) using Stat1 (D4Y6Z) Rabbit mAb #14995 (red) and DAPI #4083 (blue). The targeted mutation in the KO animal results in a reduction of expression and truncation of the protein(4). Mice from Taconic Biosciences, Inc.
Immunofluorescence Image 2: Stat1 (D4Y6Z) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed, frozen small intestine from wild type 129S6/SvEvTac mouse (model #129SVE-F) (left) or Stat1 knock-out mouse (model #2045-F) (right) using Stat1 (D4Y6Z) Rabbit mAb #14995 (red) and DAPI #4083 (blue). The targeted mutation in the KO animal results in a reduction of expression and truncation of the protein(4). Mice from Taconic Biosciences, Inc.
Immunofluorescence Image 1: Stat1 (D4Y6Z) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HeLa cells either serum-starved overnight (left) or serum-starved overnight and treated with Human Interferon-α1 (hIFN-α1) #8927 (1,000 units/ml, 30 min; right) using Stat1 (D4Y6Z) Rabbit mAb #14995 (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).
Chromatin Immunoprecipitation Image 1: Stat1 (D4Y6Z) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from HT-1080 cells treated with Human Interferon-γ (hIFN-γ) #8901 (50 ng/ml, 30 min) and either Stat1 (D4Y6Z) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human IRF-1 promoter primers, SimpleChIP® Human TAP1 Promoter Primers #5148, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.