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93130
Stat Antibody Sampler Kit II

Stat Antibody Sampler Kit II #93130

Western Blotting Image 1

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

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Western Blotting Image 2

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.

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Western Blotting Image 3

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

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Western Blotting Image 4

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

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Western Blotting Image 5

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

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Western Blotting Image 6

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

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Western Blotting Image 7

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.

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IP Image 8

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.

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IP Image 9

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.

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Western Blotting Image 10

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).

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Western Blotting Image 11

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

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Western Blotting Image 12

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.

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Chromatin IP Image 13

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Ramos cells starved overnight then treated with IL-4 (100 ng/ml, 30 min) and either 10 μl of Stat6 (D3H4) Rabbit mAb or 2 μl of 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.

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IHC-P (paraffin) Image 14

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

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Flow Cytometry Image 15

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.

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IP Image 16

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.

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Chromatin IP Image 17

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NK-92 cells starved of IL-2 overnight then treated with IL-12 (10 ng/ml) for 4 hr and either 10 μl of Stat4 (C46B10) Rabbit mAb #2653 or 2 μl of 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.

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IP Image 18

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.

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IHC-P (paraffin) Image 19

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

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IF-IC Image 20

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).

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IHC-P (paraffin) Image 21

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

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Chromatin IP Image 22

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 BaF3 cells starved of IL-3 for 6 hours followed by induction with IL-3 for 45 minutes, and either 10 μl of Stat5 (D2O6Y) Rabbit mAb or 2 μl of 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.

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Flow Cytometry Image 23

Flow cytometric analysis of ACHN cells using Stat1 (D1K9Y) 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® 488 Conjugate) #4412 was used as secondary antibody.

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Chromatin IP Image 24

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 U266 cells treated with Human Interferon-α (IFN-α) #9906 (100 ng/ml) for 30 min, and either 10 μl of Stat2 (D9J7L) Rabbit mAb or 2 μl of 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.

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IHC-P (paraffin) Image 25

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

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IF-IC Image 26

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).

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IHC-P (paraffin) Image 27

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

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Chromatin IP Image 28

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HT-1080 cells treated with Human Interferon-γ (hIFN-γ) #8901 (50 ng/ml, 30 min) and either 10 μl of Stat1 (D1K9Y) Rabbit mAb or 2 μl of 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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IHC-P (paraffin) Image 29

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

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IHC-P (paraffin) Image 30

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

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IF-IC Image 31

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).

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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
  • 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 

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.

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).

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.

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, J.E. et al. (1994) Science 264, 1415-21.
  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, 241-50.
  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.
Entrez-Gene Id
6772 , 6773 , 6774 , 6775 , 6776 , 6777 , 6778
Swiss-Prot Acc.
P42224 , P52630 , P40763 , Q14765 , P42229 , P51692 , P42226
For Research Use Only. Not For Use In Diagnostic Procedures.

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