STAT1 antibody (AA 592-731)

Catalog No. ABIN967804

Product Details anti-STAT1 Antibody

Target: STAT1 (Signal Transducer and Activator of Transcription 1, 91kDa (STAT1))

Binding Specificity: AA 592-731

Reactivity: Human, Mouse, Rat, Dog, Chicken

Host: Mouse

Clonality: Monoclonal

Conjugate: This STAT1 antibody is un-conjugated

Application: Western Blotting (WB), Immunoprecipitation (IP), BioImaging (BI)

Cross-Reactivity: Chicken, Dog (Canine), Mouse (Murine), Rat (Rattus)

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human Stat1 aa. 592-731

Clone: 42-Stat1

Isotype: IgG2b

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN968533, ABIN967389

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-STAT1 antibody (ABIN967804)

Ali, Sayeski, Bernstein: "Jak2 acts as both a STAT1 kinase and as a molecular bridge linking STAT1 to the angiotensin II AT1 receptor." in: The Journal of biological chemistry, Vol. 275, Issue 20, pp. 15586-93, (2000) (PubMed).

Zhou, Waxman: "Cross-talk between janus kinase-signal transducer and activator of transcription (JAK-STAT) and peroxisome proliferator-activated receptor-alpha (PPARalpha) signaling pathways. Growth hormone inhibition of pparalpha transcriptional activity mediated by st" in: The Journal of biological chemistry, Vol. 274, Issue 5, pp. 2672-81, (1999) (PubMed).

Rayanade, Ndubuisi, Etlinger, Sehgal: "Regulation of IL-6 signaling by p53: STAT3- and STAT5-masking in p53-Val135-containing human hepatoma Hep3B cell lines." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 161, Issue 1, pp. 325-34, (1998) (PubMed).

Akira, Nishio, Inoue, Wang, Wei, Matsusaka, Yoshida, Sudo, Naruto, Kishimoto: "Molecular cloning of APRF, a novel IFN-stimulated gene factor 3 p91-related transcription factor involved in the gp130-mediated signaling pathway." in: Cell, Vol. 77, Issue 1, pp. 63-71, (1994) (PubMed).

Ruff-Jamison, Chen, Cohen: "Induction by EGF and interferon-gamma of tyrosine phosphorylated DNA binding proteins in mouse liver nuclei." in: Science (New York, N.Y.), Vol. 261, Issue 5129, pp. 1733-6, (1993) (PubMed).

Target Details for STAT1

Target: STAT1 (Signal Transducer and Activator of Transcription 1, 91kDa (STAT1))

Alternative Name: Stat1 (STAT1 Products)

Synonyms: CANDF7 antibody, ISGF-3 antibody, STAT91 antibody, 2010005J02Rik antibody, AA408197 antibody, XStat1 antibody, STAT4 antibody, DD6G4-4 antibody, STAT antibody, STAT1 antibody, dZ182H3.3 antibody, si:by134g18.2 antibody, si:by51f19.1 antibody, si:dz182h3.3 antibody, si:dz199m19.1 antibody, si:dz73o4.2 antibody, stat1 antibody, signal transducer and activator of transcription 1 antibody, signal transducer and activator of transcription 1 L homeolog antibody, signal transducer and activator of transcription 1, 91kDa antibody, signal transducer and activator of transcription 4 antibody, signal transducer and activator of transcription 1a antibody, STAT1 antibody, Stat1 antibody, stat1.L antibody, stat4 antibody, stat1a antibody

Background: The Stat proteins function as both cytoplasmic signal transducers and activators of transcription. The Stat91/84 (the two proteins are the result of alternate splicing-Stat91 has an additional 38 C-terminal amino acids) and Stat113 were the first identified members of this protein family. These three polypeptides contain both SH2 and SH3 domains and have also been described as members of the ISGF3 (interferon-stimulated gene factor 3) complex. With the discovery of additional members of the Stat family (Stats3 & 4), the nomenclature has been revised to indicate the Stat family members in the order of their discovery. Stat 91, 84, and 113 have become Stat1alpha, Stat1beta, and Stat2, respectively. Stat1alpha is present in a higher concentration than Stat1beta in most cell types. In response to IFNalpha treatment, Stat1alpha, Stat1beta, and Stat2 become tyrosine-phosphorylated and migrate to the nucleus where they join a 48kDa DNA binding protein and subsequently direct the transcription at IFNalpha responsive elements. In IFN-gamma treated cells, Stat1alpha (but not Stat2) becomes phosphorylated and forms a dimer. It then enters the nucleus and binds to the IFN-gamma activated site (GAS) element in order to direct IFN-gamma activated transcription.

Molecular Weight: 91/84 kDa

Pathways: JAK-STAT Signaling, RTK Signaling, Interferon-gamma Pathway, Response to Growth Hormone Stimulus, Cellular Response to Molecule of Bacterial Origin, Positive Regulation of Endopeptidase Activity, Hepatitis C, CXCR4-mediated Signaling Events