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93130
Stat Antibody Sampler Kit II
Primary Antibodies
Antibody Sampler Kit

Stat Antibody Sampler Kit II #93130

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Stat Antibody Sampler Kit II: Image 1

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.

Stat Antibody Sampler Kit II: Image 2

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.

Stat Antibody Sampler Kit II: Image 3

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 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #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 data sheet.

Stat Antibody Sampler Kit II: Image 4

Confocal immunofluorescent analysis of HeLa cells (Stat3 positive), serum-starved (left) or treated with Human Interleukin-6 (hIL-6) #8904 (100 ng/mL, 20 min; center), or PC-3 cells (Stat3 negative) treated with Human Interleukin-6 (hIL-6) #8904 (100 ng/mL, 20 min; right), using Stat3 (D1B2J) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).

Stat Antibody Sampler Kit II: Image 5

Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells starved of IL-2 overnight then treated with IL-12 (10 ng/ml) for 4 hr and Stat4 (C46B10) 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 IRF1, a known target gene of Stat4 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

Stat Antibody Sampler Kit II: Image 6

Chromatin immunoprecipitations were performed with cross-linked chromatin from BaF3 cells starved of IL-3 for 6 hours followed by induction with IL-3 for 45 minutes, and either Stat5 (D2O6Y) 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® Mouse CIS Intron 1 Primers #5131, mouse SOCS-3 promoter primers, and SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. 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.

Stat Antibody Sampler Kit II: Image 7

Chromatin immunoprecipitations were performed with cross-linked chromatin from Ramos cells starved overnight then treated with IL-4 (100 ng/ml, 30 min) and either Stat6 (D3H4) 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 DMD Intron 2 Primers #7710, human MS4A1 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.

Stat Antibody Sampler Kit II: Image 8

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.

Stat Antibody Sampler Kit II: Image 9

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.

Stat Antibody Sampler Kit II: Image 10

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 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #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).

Stat Antibody Sampler Kit II: Image 11

Immunohistochemical analysis of paraffin-embedded HeLa cell pellet (left, positive) or PC-3 cell pellet (right, negative) using Stat3 (D1B2J) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 12

Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells starved of IL-2 overnight then treated with IL-12 (10 ng/ml) for 4 hr and Stat4 (C46B10) 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 chromosome 5 (upper), including IRF1 (lower), a known target gene of Stat4 (see additional figure containing ChIP-qPCR data).

Stat Antibody Sampler Kit II: Image 13

Immunoprecipitation of Stat5 from K-562 cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Stat5 (D2O6Y) Rabbit mAb. Western blot was performed using Stat5 (D2O6Y) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 14

Western blot analysis of extracts from various cell lines using Stat6 (D3H4) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 15

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).

Stat Antibody Sampler Kit II: Image 16

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.

Stat Antibody Sampler Kit II: Image 17

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Stat3 (D1B2J) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 18

Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells starved of IL-2 overnight then treated with IL-12 (10 ng/ml) for 4 hr and either Stat4 (C46B10) Rabbit mAb #2653 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, human PRF1 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.

Stat Antibody Sampler Kit II: Image 19

Western blot analysis of extracts from various cell lines using Stat5 (D2O6Y) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Stat Antibody Sampler Kit II: Image 20

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Stat1 (D1K9Y) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 21

Flow cytometric analysis of U266 cells using Stat2 (D9J7L) Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 647 Conjugate) was used as a secondary antibody.

Stat Antibody Sampler Kit II: Image 22

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Stat3 (D1B2J) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 23

Immunoprecipitation of Stat4 protein from NK-92 cell extracts. Lane 1 is 10% input, lane 2 is beads only, lane 3 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 4 is Stat4 (C46B10) Rabbit mAb. Western blot analysis was performed using Stat4 (C46B10) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 24

Western blot analysis of PC-3 cells, mock transfected (-) or transfected with constructs expressing full-length human Stat5a (+) or Stat5b (+) using Stat5 (D2O6Y) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 25

Immunohistochemical analysis of paraffin-embedded human lymphoma using Stat1 (D1K9Y) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 26

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).

Stat Antibody Sampler Kit II: Image 27

Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Stat3 (D1B2J) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 28

Western blot analysis of extracts from various cell lines and from rat thymus using Stat4 (C46B10) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 29

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.

Stat Antibody Sampler Kit II: Image 30

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.

Stat Antibody Sampler Kit II: Image 31

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Stat3 (D1B2J) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 32

Western blot analysis of extracts from ACHN, SR and Caki cell lines and from rat thymus using Stat4 (C46B10) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 33

Western blot analysis of extracts from various cell lines using Stat1 (D1K9Y) Rabbit mAb.

Stat Antibody Sampler Kit II: Image 34

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.

Stat Antibody Sampler Kit II: Image 35

Immunoprecipitation of Stat3 from MCF7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Stat3 (D1B2J) Rabbit mAb. Western blot was performed using Stat3 (D1B2J) Rabbit mAb. A conformation-specific secondary antibody was used to avoid reactivity with IgG.

Stat Antibody Sampler Kit II: Image 36

Western blot analysis of extracts from various cell lines using Stat3 (D1B2J) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Stat Antibody Sampler Kit II: Image 37

Western blot analysis of extracts from NIH/3T3 cells, mock transfected (-) or transfected with an siRNA against mouse Stat3 (mStat3 siRNA; +), using Stat3 (D1B2J) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

To Purchase # 93130T
Product # Size Price
93130T
1 Kit  (6 x 20 µl) $ 445

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Stat1 (D1K9Y) Rabbit mAb 14994 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Mk 84, 91 Rabbit IgG
Stat2 (D9J7L) Rabbit mAb 72604 20 µl
  • WB
  • IP
  • IF
  • F
  • ChIP
H M 97, 113 Rabbit IgG
Stat3 (D1B2J) Rabbit mAb 30835 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R 79, 86 Rabbit IgG
Stat4 (C46B10) Rabbit mAb 2653 20 µl
  • WB
  • IP
  • ChIP
H M R 81 Rabbit 
Stat5 (D2O6Y) Rabbit mAb 94205 20 µl
  • WB
  • IP
  • ChIP
H M R 90 Rabbit IgG
Stat6 (D3H4) Rabbit mAb 5397 20 µl
  • WB
  • IP
  • ChIP
H M R 110 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

Stat Antibody Sampler Kit II provides an economical means to examine the complete Stat family: Stat1-6. The kit contains enough a primary antibody to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each of the antibodies in the Stat Antibody Sampler Kit II recognizes endogenous levels of their target proteins. Stat1 (D1K9Y) Rabbit mAb recongizes an unidentified protein at 150 kDa. Stat3 (D1B2J) Rabbit mAb has some unclear staining observed in rodent. Stat5 (D2O6Y) Rabbit mAb recognizes both alpha and beta isoforms of Stat5 (Stat5a and Stat5b).

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro688 of human Stat1 protein, Leu706 of human Stat2, Pro695 of human Stat3, Lys151 of human Stat4, Leu60 of human Stat5, and Gly582 of human Stat6.

Background

Jaks (Janus Kinases) and Stats (Signal Transducers and Activators of Transcription) are utilized by receptors for a wide variety of ligands including cytokines, hormones, growth factors and neurotransmitters. Jaks, activated via autophosphorylation following ligand-induced receptor aggregation, phosphorylate tyrosine residues on associated receptors, Stat molecules and other downstream signaling proteins (1,2). The phosphorylation of Stat proteins at conserved tyrosine residues activates SH2-mediated dimerization followed rapidly by nuclear translocation. Stat dimers bind to IRE (interferon response element) and GAS (gamma interferon-activated sequence) DNA elements, resulting in the transcriptional regulation of downstream genes (1,2). The remarkable range and specificity of responses regulated by the Stats is determined in part by the tissue-specific expression of different cytokine receptors, Jaks and Stats (2,3), and by the combinatorial coupling of various Stat members to different receptors. Serine phosphorylation in the carboxy-terminal transcriptional activation domain has been shown to regulate the function of Stat1, -2, -3, -4 and -5 (1). Phosphorylation of Stat3 at Ser727 via MAPK or mTOR pathways is required for optimal transcriptional activation in response to growth factors and cytokines including IFN-gamma and CNTF (4,5). Jak/Stat pathways also play important roles in oncogenesis, tumor progression, angiogenesis, cell motility, immune responses and stem cell differentiation (6-11).

  1. Darnell Jr., J. et al. (1994) Science 264, 1415-1421.
  2. Leonard, W.J. and O'Shea, J.J. (1998) Annu. Rev. Immunol. 16, 293-322.
  3. Caldenhoven, E. et al. (1996) J. Biol. Chem. 271, 13221-13227.
  4. Wen, Z. et al. (1995) Cell 82, 241-250.
  5. Yokogami, K. et al. (2000) Curr. Biol. 10, 47-50.
  6. Lim, C.P. and Cao, X. (1999) J. Biol. Chem. 274, 31055-31061.
  7. Bromberg, J. F. et al. (1999) Cell 98, 295-303.
  8. Su, L. et al. (1999) J. Biol. Chem. 274, 31770-31774.
  9. Dentelli, P. et al. (1999) J. Immunol. 163, 2151-2159.
  10. Cattaneo, E. et al. (1999) Trends Neurosci. 22, 365-369.
  11. Frank, D.A. (1999) Mol. Med. 5, 432-456.

Pathways & Proteins

Explore pathways + proteins related to this product.

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U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.