Revision 1

#35114Store at -20C

1 Kit

(9 x 20 microliters)

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.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Tyk2 (D4I5T) Rabbit mAb 14193 20 µl 134 kDa Rabbit IgG
Phospho-Tyk2 (Tyr1054/1055) (D7T8A) Rabbit mAb 68790 20 µl 134 kDa Rabbit IgG
Jak1 (6G4) Rabbit mAb 3344 20 µl 130 kDa Rabbit IgG
Phospho-Jak1(Tyr1034/1035) (D7N4Z) Rabbit mAb 74129 20 µl 130 kDa Rabbit IgG
Stat1 (D1K9Y) Rabbit mAb 14994 20 µl 84, 91 kDa Rabbit IgG
Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb 7649 20 µl 84, 91 kDa Rabbit IgG
Stat2 (D9J7L) Rabbit mAb 72604 20 µl 97, 113 kDa Rabbit IgG
Phospho-Stat2 (Tyr690) (D3P2P) Rabbit mAb 88410 20 µl 97, 113 kDa Rabbit IgG
IRF-9 (D2T8M) Rabbit mAb 76684 20 µl 48 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The IFN (Type I/III) Signaling Pathway Antibody Sampler Kit provides an economical means of detecting the activation of the IFN (Type I/III) signaling pathway using phospho-specific and control antibodies. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

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 antibodies.

Background

Originally discovered in the late 1950s for their antiviral activity, interferons (IFNs) have since been assigned diverse roles in many physiological and pathological processes. There are three families of IFNs: types I, II, and III. In humans, type I contains IFN-α (13 different subtypes), IFN-β (also known as IFN-β1), IFN-ε, IFN-κ, and IFN-ω. They bind to a receptor complex containing IFNAR1 and IFNAR2, which is broadly expressed on most cells. IFN-γ is the sole member of type II IFN. It signals through a receptor complex consisting of IFNγR1 and IFNγR2, which is also expressed on most cell types. Type III IFN, also known as interferon lambdas (IFN-λs), have four members in humans: IFN-λ1 (IL29), IFN-λ2 (IL28A), IFN-λ3 (IL28B), and IFN-λ4. IFN-λs signal through a heterodimeric receptor comprised of IFNλR1 and IL-10R2. While IL-10R2 is broadly expressed and shared by the IL-10 family cytokines, IFNλR1 expression is restricted to epithelial cells, neuronal cells, and subsets of myeloid cells (1-3). Engagement of all IFNs with their receptors initiates downstream signaling events, mainly, activation of the Jak–Stat signaling cascade. For type I and III IFNs, Jak1 and Tyk2 are phosphorylated and activated, leading to subsequent phosphorylation of Stat1 and Stat2. Phosphorylated Stat1 and Stat2 are released from the receptor complex and, together with IRF-9, they form so-called ISGF3 (interferon-stimulated gene factor 3) transcriptional complex. ISGF3 translocates to the nucleus, binds to the interferon-stimulated response element (ISRE) to initiate the transcription of a wide array of interferon-stimulated genes (ISGs) (4,5). On the other hand, IFN-γ induces phosphorylation and activation of Jak1 and Jak2, which subsequently phosphorylate Stat1. Phosphorylated Stat1 dimerizes, translocates to the nucleus, and binds to γ-interferon-activated site (GAS) to initiate the transcription of ISGs (6,7).

  1. Schneider, W.M. et al. (2014) Annu Rev Immunol 32, 513-45.
  2. Hemann, E.A. et al. (2017) Front Immunol 8, 1707.
  3. Walter, M.R. (2020) Front Immunol 11, 606489.
  4. Hervas-Stubbs, S. et al. (2011) Clin Cancer Res 17, 2619-27.
  5. Mesev, E.V. et al. (2019) Nat Microbiol 4, 914-924.
  6. Green, D.S. et al. (2017) J Biol Chem 292, 13925-13933.
  7. Ivashkiv, L.B. (2018) Nat Rev Immunol 18, 545-558.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    Jak antibodies produced under license (granting certain rights including those under U.S. Patent No. 5,658,791) from Chemicon International, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    For Research Use Only. Not for Use in Diagnostic Procedures.
    Revision 1
    #35114

    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit

    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 1 Expand Image
    CUT&RUN was performed with HT-1080 cells treated with (hIFN-γ) #8901 (50 ng/ml, 30 min) and Stat1 (D1K9Y) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human AIM2 promoter primers, human FZD5 promoter primers, 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.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 2 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from MCF-7 cells using Stat1 (D1K9Y) Rabbit mAb #14994. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 3 Expand Image
    CUT&RUN was performed with HT-1080 cells and Stat1 (D1K9Y) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across FZD5 gene, a known target gene of Stat1 (see additional figure containing CUT&RUN-qPCR data).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 4 Expand Image
    CUT&RUN was performed with HT-1080 cells and Stat1 (D1K9Y) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 2 (upper), including FZD5 gene (lower), a known target gene of Stat1 (see additional figure containing CUT&RUN-qPCR data).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 5 Expand Image
    Flow cytometric analysis of K-562 cells using Stat2 (D9J7L) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from various cell lines using Tyk2 (D4I5T) Rabbit mAb.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from A549 cells (lane 1) or STAT1 knock-out cells (lane 2) using Stat1 (D1K9Y) Rabbit mAb #14994 (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.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from ACHN, SR, CTLL-2 and PC12 cell lines using Jak1 (6G4) Rabbit mAb.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from serum-starved U266 and Jurkat cells, untreated (-) or treated with Human Interferon-α1 (hIFN-α1) #8927 (50 ng/ml, 15 min; +) using Phospho-Tyk2 (Tyr1054/1055) (D7T8A) Rabbit mAb (upper) or total Tyk2 (D4I5T) Rabbit mAb #14193 (lower).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 10 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from various cell lines using Stat2 (D9J7L) Rabbit mAb. KARPAS cell line source: Dr Abraham Karpas at the University of Cambridge.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 12 Expand Image
    CUT&RUN was performed with U266 cells treated with Human Interferon-α1 (hIFN-α1) #8927 (10nM, 30min) and Stat2 (D9J7L) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across GAS5 gene.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from various cell lines, serum-starved overnight (-) followed by treatment with Human Interferon-α1 #8927 (hIFN-α1, 10 ng/ml, 15 min; +) or Human Interleukin-4 #8919 (hIL-4, 10 ng/ml, 10 min; +) using Phospho-Jak1 (Tyr1034/1035) (D7N4Z) Rabbit mAb (upper) or Jak1 (6G4) Rabbit mAb (lower).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from HeLa, A20, and PC-12 cells, untreated or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, 30 min), using Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb (upper) or Stat1 Antibody #9172 (lower).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 15 Expand Image
    Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, 16 hr; +), using IRF-9 (D2T8M) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 16 Expand Image
    Western blot analysis of extracts from serum-starved U266 and A-431 cells, untreated (-) or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml; +) using Phospho-Stat2 (Tyr690) (D3P2P) Rabbit mAb (upper) and total Stat2 (D9J7L) Rabbit mAb #72604 (lower).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 17 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human Tyk2 (hTyk2; +), using Tyk2 (D4I5T) Rabbit mAb.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 18 Expand Image
    Western blot analysis of extracts from various cell lines using Stat1 (D1K9Y) Rabbit mAb.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Jak1 (6G4) Rabbit mAb #3344, in the presence of control peptide (left) or Jak1 Blocking Peptide (right).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 20 Expand Image
    Immunoprecipitation of Stat2 from KARPAS-299 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Stat2 (D9J7L) Rabbit mAb. Western blot was performed using Stat2 (D9J7L) Rabbit mAb. KARPAS cell line source: Dr Abraham Karpas at the University of Cambridge.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 21 Expand Image
    CUT&RUN was performed with U266 cells treated with Human Interferon-α1 (hIFN-α1) #8927 (10nM, 30min) and Stat2 (D9J7L) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 1 (upper), including GAS5 gene (lower).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 22 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with hIFN-α1 #8927 (100 ng/mL, 30 min; right), using Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 23 Expand Image
    Western blot analysis of extracts from various cell lines using IRF-9 (D2T8M) Rabbit mAb.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 24 Expand Image
    Confocal immunofluorescent analysis of A-431 cells, serum starved (left) or treated with IFNα (1000 U/ml for 30 min; right) using Phospho-Stat2 (Tyr690) (D3P2P) Rabbit mAb (green) and β-actin (8H10D10) Mouse mAb #3700 (red).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 25 Expand Image
    Western blot analysis of extracts from MCF7 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Tyk2 siRNA #14254 (+), using Tyk2 (D4I5T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The Tyk2 (D4I5T) Rabbit mAb confirms silencing of Tyk2 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 26 Expand Image
    Immunoprecipitation of Stat1 from MCF7 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Stat1 (D1K9Y) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using Stat1 (9H2) Mouse mAb #9176.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Jak1 (6G4) Rabbit mAb.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 28 Expand Image
    Confocal immunofluorescent analysis of A-431 cells, serum starved (left) or treated with IFNα (1000 U/ml for 30 mins; right) using Stat2 (D9J7L) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 29 Expand Image
    CUT&RUN was performed with U266 cells treated with Human Interferon-α1 (hIFN-α1) #8927 (10nM, 30min) and either Stat2 (D9J7L) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human SIN3A promoter primers and human ITM2A upstream primers. 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.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 30 Expand Image
    Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with hIFN-α1 #8927 (green), using Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 31 Expand Image
    Immunoprecipitation of IRF-9 from THP-1 cell extracts. Lane 1 is 10% input, lane 2 is with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is with IRF-9 (D2T8M) Rabbit mAb. Western blot analysis was performed using IRF-9 (D2T8M) Rabbit mAb.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 32 Expand Image
    Flow cytometric analysis of U266 cells, untreated (blue) or treated with IFN-α (green) using Phospho-Stat2 (Tyr690) (D3P2P) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary antibody.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 33 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Stat1 (D1K9Y) Rabbit mAb.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Jak1 (6G4) Rabbit mAb.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 35 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across IRF-1, a known target gene of Phospho-Stat1 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 36 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, 16 hr; right), using IRF-9 (D2T8M) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 37 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lymphoma using Stat1 (D1K9Y) Rabbit mAb.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 38 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from U266 cells treated with Human Interferon-α (IFN-α) #9906 (10nM) for 30 min and Stat2 (D9J7L) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across USP18, a known target gene of Stat2 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 39 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and 5 μl of Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 5 (upper), including IRF1 (lower), a known target gene of Phospho-Stat1 (see additional figure containing ChIP-qPCR data).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 40 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from U266 cells treated with Human Interferon-α1 (hIFN-α1) #8927 (10 nM, 30 min) and either IRF-9 (D2T8M) 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 SimpleChIP® Human MX1 Promoter Primers #57949, human WARS intron 1 primers, 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.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 41 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, serum-starved overnight (left) or treated with Human Interferon-α1 (hIFN-α1) #8927 (1,000 units/ml, 30 min; right), using Stat1 (D1K9Y) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 42 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from U266 cells treated with Human Interferon-α (IFN-α) #9906 (10nM) for 30 min and Stat2 (D9J7L) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 22 (upper), including USP18 (lower), a known target gene of Stat2 (see additional figure containing ChIP-qPCR data).
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 43 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and either Phospho-Stat1 (Tyr701) (D4A7) 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.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 44 Expand Image
    Flow cytometric analysis of ACHN cells using Stat1 (D1K9Y) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 45 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from U266 cells treated with Human Interferon-α (IFN-α) #9906 (100 ng/ml) for 30 min, and either Stat2 (D9J7L) 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 USP18 promoter primers, SimpleChIP® Human WARS Intron 1 Primers #30101, 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.
    IFN (Type I/III) Signaling Pathway Antibody Sampler Kit: Image 46 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 (D1K9Y) 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.
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    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    For Research Use Only. Not for Use in Diagnostic Procedures.