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57103
IRF Family Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

IRF Family Antibody Sampler Kit #57103

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Flow cytometric analysis of HeLa cells, untreated (blue) or treated with Human Interferon-γ (hIFN-γ) #8901 (green), using IRF-1 (D5E4) XP® Rabbit mAb.

Western blot analysis of extracts from SR, Ramos and A20 cells using IRF-2 Antibody.

Western blot analysis of extracts from control HeLa cells (lane 1) or IRF-3 knockout HeLa cells (lane 2) using IRF-3 (D6I4C) XP® Rabbit mAb, #11904 (upper) or β-actin (13E5) Rabbit mAb, #4970 (lower). The absence of signal in the IRF-3 knockout HeLa cells confirms specificity of the antibody for IRF-3.

Confocal immunofluorescent analysis of HT-29 cells, untransfected (left) or transfected with Poly(I:C) (2.5 μg/ml, 6 hr; right), using IRF-3 (D6I4C) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Chromatin immunoprecipitations were performed with cross-linked chromatin from H929 cells and either IRF-4 (D9P5H) 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 PRDM1 Exon 2 primers, SimpleChIP® Human SUB1 Promoter Primers #5156 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.

Chromatin immunoprecipitations were performed with cross-linked chromatin from H929 cells and IRF-4 (D9P5H) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across SSR2 gene. For additional ChIP-seq tracks, please download the product data sheet.

Immunoprecipitation of IRF-5 from HDLM-2 cell extracts. Lane 1 is 10% input, lane 2 is Normal Rabbit IgG #2729, and lane 3 is IRF-5 Antibody. Western blot analysis was performed using IRF-5 Antibody. Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 was used for detection.

Western blot analysis of extracts from various cell lines using IRF-6 Antibody. Absence of IRF-6 in MCF7 cells has been reported previously (5). Doublet has been reported to result from phosphorylation (6).

Western blot analysis of extracts from HT-29 and G-361 cells, untreated (-) or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, overnight; +), using IRF-7 (D2A1J) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Chromatin immunoprecipitations were performed with cross-linked chromatin from THP-1 and IRF-8 (D20D8) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across GPR18/UBAC2, a known target gene of IRF8 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

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.

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).

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.

Confocal immunofluorescent analysis of HeLa cells, serum-starved (left) or treated with hIFNγ #8901 (100 ng/mL, 4 hr; right), using IRF-1 (D5E4) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin.

Immunoprecipitation of IRF-3 from THP-1 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or IRF-3 (D6I4C) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using IRF-3 (D6I4C) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 and Anti-mouse IgG, HRP-linked Antibody #7076 were used as secondary antibodies.

Western blot analysis of extracts from various cell lines using IRF-5 Antibody (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western blot analysis of extracts from HT-29 cells transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® IRF-7 siRNA I #13139 (+) or SignalSilence® IRF-7 siRNA II #13291 (+). Twenty-four hours after transfection, cells were treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, overnight; +) and analyzed by western blot using IRF-7 (D2A1J) Rabbit mAb #13014 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The IRF-7 (D2A1J) Rabbit mAb confirms silencing of IRF-7 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 THP-1 and either IRF-8 (D20D8) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using human GPR18 promoter primers, SimpleChIP® Human MS4A7 Promoter Primers #9013, 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.

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).

Imunohistochemical analysis of paraffin-embedded human breast carcinoma using IRF-1 (D5E4) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Western blot analysis of extracts from various cell lines using IRF-3 (D6I4C) XP® Rabbit mAb.

Immunoprecipitation of IRF-4 from RPMI 8226 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is IRF-4 (D9P5H) Rabbit mAb. Western blot analysis was performed using IRF-4 (D9P5H) Rabbit mAb.

Western blot analysis of extracts from Raji and Jurkat cells, untreated or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml; overnight) using IRF-8 (D20D8) Rabbit mAb.

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.

Imunohistochemical analysis of paraffin-embedded human lymphoma using IRF-1 (D5E4) XP® Rabbit mAb.

Western blot analysis of extracts from various cell lines using IRF-4 (D9P5H) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western blot analysis of extracts from THP-1, RL7, and A20 cell lines using IRF-8 (D20D8) Rabbit mAb.

Western blot analysis of extracts from various cell lines using IRF-9 (D2T8M) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded mosue thymus using IRF-1 (D5E4) XP® Rabbit mAb.

Western blot analysis of extracts from 293T cells, mock transfected or transfected with human and mouse IRF-8 constructs, using IRF-8 (D20D8) Rabbit mAb.

Imunohistochemical analysis of paraffin-embedded HeLa cell pellets, control (left) or IFN gamma-treated (right), using IRF-1 (D5E4) XP® Rabbit mAb.

Western blot analysis of HeLa cells, untreated or treated with Human Interferon-γ (hIFN-γ) #8901 (100 ng/ml, 4 hr), using IRF-1 (D5E4) XP® Rabbit mAb.

Western blot analysis of extracts from various cell lines using IRF-1 (D5E4) XP® Rabbit mAb.

To Purchase # 57103T
Product # Size Price
57103T
1 Kit  (9 x 20 µl) $ 609

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
IRF-1 (D5E4) XP® Rabbit mAb 8478 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R 45-48 Rabbit IgG
IRF-2 Antibody 4943 20 µl
  • WB
  • IP
H M R 45 Rabbit 
IRF-3 (D6I4C) XP® Rabbit mAb 11904 20 µl
  • WB
  • IP
  • IF
H Mk 50-55 Rabbit IgG
IRF-4 (D9P5H) Rabbit mAb 15106 20 µl
  • WB
  • IP
  • ChIP
H M R 51 Rabbit IgG
IRF-5 Antibody 20261 20 µl
  • WB
  • IP
H M 60 Rabbit 
IRF-6 Antibody 6948 20 µl
  • WB
  • IP
H 58, 60 Rabbit 
IRF-7 (D2A1J) Rabbit mAb 13014 20 µl
  • WB
H 65 Rabbit IgG
IRF-8 (D20D8) Rabbit mAb 5628 20 µl
  • WB
  • IP
  • ChIP
H M 50 Rabbit IgG
IRF-9 (D2T8M) Rabbit mAb 76684 20 µl
  • WB
  • IP
  • IF
  • ChIP
H 48 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The IRF Family Antibody Sampler Kit contains reagents to examine the total protein levels of various IRF isoforms. The kit contains enough primary and secondary antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the IRF Family Antibody Sampler Kit detects endogenous levels of its target protein. IRF-8 (D20D8) Rabbit mAb also detects an unknown background band at 80 kDa in some cell lines. IRF-9 (D2T8M) Rabbit mAb cross-reacts with an unidentified protein at 95 kDa.

Source / Purification

Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to residues surrounding Pro261 of human IRF-1, Pro115 of human IRF-7, and Gly65 of human IRF-8, or recombinant human IRF-3, mouse IRF-4, and human IRF-9 proteins. Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Asp327 of human IRF-2, Gly121 of human IRF-6, and near the carboxy terminus of human IRF-5. Antibodies were purified by affinity chromatography.

Background

Interferon regulatory factors (IRFs) comprise a family of transcription factors that function within the Jak/Stat pathway to regulate interferon (IFN) and IFN-inducible gene expression in response to viral infection (1). IRFs play an important role in pathogen defense, autoimmunity, lymphocyte development, cell growth, and susceptibility to transformation. The IRF family includes nine members: IRF-1, IRF-2, IRF-9/ISGF3γ, IRF-3, IRF-4 (Pip/LSIRF/ICSAT), IRF-5, IRF-6, IRF-7, and IRF-8/ICSBP. All IRF proteins share homology in their amino-terminal DNA-binding domains. IRF family members regulate transcription through interactions with proteins that share similar DNA-binding motifs, such as IFN-stimulated response elements (ISRE), IFN consensus sequences (ICS), and IFN regulatory elements (IRF-E) (2).

  1. Taniguchi, T. et al. (2001) Annu Rev Immunol 19, 623-55.
  2. Honda, K. and Taniguchi, T. (2006) Nat Rev Immunol 6, 644-58.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.

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