Microsize antibodies for $99 | Learn More >>
19171
Microglia Interferon-Related Module Antibody Sampler Kit
Primary Antibodies

Microglia Interferon-Related Module Antibody Sampler Kit #19171

Western Blotting Image 1

Western blot analysis of extracts from J774A.1 and Raw 264.7 cells using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Learn more about how we get our images
Western Blotting Image 2

Western blot analysis of cell extracts from Baf3, 32D, and mouse spleen using HS1 (D5A9) XP® Rabbit mAb (Rodent Specific).

Learn more about how we get our images
Chromatin IP-seq 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.

Learn more about how we get our images
Western Blotting Image 4

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.

Learn more about how we get our images
Western Blotting Image 5

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).

Learn more about how we get our images
Western Blotting Image 6

Western blot analysis of extracts from various cell lines and tissues using Akt3 (E1Z3W) Rabbit mAb (upper) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).

Learn more about how we get our images
Western Blotting Image 7

Western blot analysis of extracts from PC-3 cells, untreated or LY294002/wortmannin-treated, and NIH/3T3 cells, serum-starved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).

Learn more about how we get our images
Western Blotting 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.

Learn more about how we get our images
IP Image 9

Immunoprecipitation of ASC/TMS1 from J774A.1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is ASC (D2W8U) Rabbit mAb (Mouse Specific). Western blot analysis was performed using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific).

Learn more about how we get our images
IHC-P (paraffin) Image 10

Immunohistochemical analysis of paraffin-embedded mouse spleen using HS1 (D5A9) XP® Rabbit mAb (Rodent Specific).

Learn more about how we get our images
Chromatin IP Image 11

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.

Learn more about how we get our images
IP Image 12

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.

Learn more about how we get our images
Flow Cytometry Image 13

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.

Learn more about how we get our images
Western Blotting Image 14

Western blot analysis of recombinant Akt1, Akt2, and Akt3 proteins using Akt3 (E1Z3W) Rabbit mAb (upper) and Akt1 (pan) (C67E7) Rabbit mAb #4691 (lower).

Learn more about how we get our images
IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded MDA-MB-468 xenograft using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (left) or PTEN (138G6) Rabbit mAb #9559 (right). Note the presence of P-Akt staining in the PTEN deficient MDA-MB-468 cells.

Learn more about how we get our images
Flow Cytometry Image 16

Flow cytometric analysis of Raw264.7 cells (blue) and J774A.1 cells (green) using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific) (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Learn more about how we get our images
IHC-P (paraffin) Image 17

Immunohistochemical analysis of paraffin-embedded LL2 syngeneic tumor using HS1 (D5A9) XP® Rabbit mAb (Rodent Specific).

Learn more about how we get our images
Flow Cytometry Image 18

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.

Learn more about how we get our images
IF-IC Image 19

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).

Learn more about how we get our images
IF-IC Image 20

Confocal immunofluorescent analysis of A172 (positive, left) or LNCaP (negative, right) cells using Akt3 (E1Z3W) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Learn more about how we get our images
IHC-P (paraffin) Image 21

Immunohistochemical analysis of paraffin-embedded human breast carcinoma comparing SignalStain® Antibody Diluent #8112 (left) to TBST/5% normal goat serum (right) using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060.

Learn more about how we get our images
IF-IC Image 22

Confocal immunofluorescent analysis of mouse primary bone marrow-derived macrophages (BMDMs) either untreated (upper left) or treated with LPS (50 ng/ml, 4 hr, middle) or LPS followed by ATP (5 mM, 45 min, upper right), and J774A.1 (lower left) or Raw 264.7 (lower right) cells, using ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Note the translocation of ASC to inflammasomes following stimulation with LPS and ATP (white arrows).

Learn more about how we get our images
Flow Cytometry Image 23

Flow cytometric analysis of NIH/3T3 cells (red) and 32D cells (blue), using HS1 (D5A9) XP® Rabbit mAb (Rodent Specific).

Learn more about how we get our images
IF-IC Image 24

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).

Learn more about how we get our images
IHC-P (paraffin) Image 25

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.

Learn more about how we get our images
IF-IC Image 26

Confocal immunofluorescent analysis of 32D cells (left) and C2C12 cells (right), using HS1 (D5A9) XP® Rabbit mAb (Rodent Specific) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Learn more about how we get our images
IHC-P (paraffin) Image 27

Immunohistochemical analysis using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb on SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)).

Learn more about how we get our images
IF-F Image 28

Confocal immunofluorescent analysis of mouse Tg2576 brain which overexpresses mutant human APP695. Sections were first labeled with HS1 (D5A9) XP® Rabbit mAb (Rodent Specific) (green) and APP/β-Amyloid (NAB228) Mouse mAb #2450 (yellow). After blocking free secondary binding sites with Mouse (G3A1) mAb IgG1 Isotype Control #5415, sections were incubated with GFAP (GA5) Mouse mAb (Alexa Fluor® 647 Conjugate) #3657 (red). Nuclei were labeled with Hoechst 33342 #4082 (blue).

Learn more about how we get our images
IHC-P (paraffin) Image 29

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 30

Immunohistochemical analysis of paraffin-embedded PTEN heterozygous mutant mouse endometrium using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb. (Tissue section courtesy of Dr. Sabina Signoretti, Brigham and Women's Hospital, Harvard Medical School, Boston, MA.)

Learn more about how we get our images
IHC-P (paraffin) Image 31

Immunohistochemical analysis of paraffin-embedded U-87MG xenograft, untreated (left) or lambda phosphatase-treated (right), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.

Learn more about how we get our images
Flow Cytometry Image 32

Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, wortmannin #9951 and U0126 #9903 (blue), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb compared to a nonspecific negative control antibody (red).

Learn more about how we get our images
IF-IC Image 33

Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin #8953 (red). Blue pseudocolor = DRAQ5®#4084 (fluorescent DNA dye).

Learn more about how we get our images
Product Includes Quantity Applications Reactivity MW(kDa) Isotype
ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific) 67824 20 µl
  • WB
  • IP
  • IF
  • F
M 22 Rabbit IgG
HS1 (D5A9) XP® Rabbit mAb (Rodent Specific) 3892 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
M R 80 Rabbit IgG
Stat2 (D9J7L) Rabbit mAb 72604 20 µl
  • WB
  • IP
  • IF
  • F
  • ChIP
H M 97, 113 Rabbit IgG
Phospho-Stat2 (Tyr690) (D3P2P) Rabbit mAb 88410 20 µl
  • WB
  • IF
  • F
H R 97, 113 Rabbit IgG
Akt3 (E1Z3W) Rabbit mAb 14982 20 µl
  • WB
  • IP
  • IF
H M R 60 Rabbit IgG
Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb 4060 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Hm Mk Dm Z B 60 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Microglia Interferon-Related Module Antibody Sampler Kit provides an economical means of detecting proteins identified as markers of interferon-related microglial activity by western blot and/or immunofluorescence.

Each antibody in the Microglia Interferon-Related Module Antibody Sampler Kit detects endogenous levels of its target protein. Phospho-Stat2 (Tyr690) (D3P2P) Rabbit mAb recognizes endogenous levels of Stat2 protein only when phosphorylated at Tyr690. Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb detects endogenous levels of Akt only when phosphorylated at Ser473. HS1 (D5A9) XP® Rabbit mAb (Rodent Specific) does not recognize human HS1 protein. HS1 has a calculated size of 54 kDa, but has an apparent molecular weight of 80 kDa on SDS-PAGE gels.

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu310 of mouse HS1, His140 of human Akt3, Leu706 of human Stat2, a phospho-specific synthetic peptide corresponding to residues surrounding Tyr690 of human Stat2 protein and Ser473 of human Akt, and recombinant mouse ASC/TMS1 protein.

Distinct microglial activation states have been identified using RNA-seq data from a vast array of neurological disease and aging models. These activation states have been categorized into modules corresponding to proliferation, neurodegeneration, interferon-relation, LPS-relation, and many others (1). Previous work identifying markers of specific brain cell types using RNA-seq has shown HS1 and ASC/TMS1 to be useful and specific tools to study microglia (2). HS1 is a protein kinase substrate that is expressed only in tissues and cells of hematopoietic origin (3) and ASC/TMS1 has been found to be a critical component of inflammatory signaling where it associates with and activates caspase-1 in response to pro-inflammatory signals (4).

Stat2 is critical to the transcriptional responses induced by type I interferons, IFN-alpha/beta (5,6). In response to IFN-alpha/beta, Stat2 is activated by phosphorylation at site Tyr690 through associations with receptor-bound Jak kinases (7). Akt is a protein kinase that plays a critical role in controlling survival and apoptosis. Akt is activated by various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (8-10) and its activity is shown to be essential for up-regulation of key IFN inducible proteins (11). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (12) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (13,14).

  1. Friedman, B.A. et al. (2018) Cell Rep 22, 832-47.
  2. Zhang, Y. et al. (2014) J Neurosci 34, 11929-47.
  3. Kitamura, D. et al. (1995) Biochem Biophys Res Commun 208, 1137-46.
  4. Srinivasula, S.M. et al. (2002) J Biol Chem 277, 21119-22.
  5. Kaur, S. et al. (2008) Proc Natl Acad Sci U S A 105, 4808-13.
  6. Fu, X.Y. et al. (1992) Proc Natl Acad Sci U S A 89, 7840-3.
  7. Ihle, J.N. (2001) Curr Opin Cell Biol 13, 211-7.
  8. Franke, T.F. et al. (1997) Cell 88, 435-7.
  9. Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.
  10. Franke, T.F. et al. (1995) Cell 81, 727-36.
  11. Improta, T. et al. (1994) Proc Natl Acad Sci U S A 91, 4776-80.
  12. Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
  13. Sarbassov, D.D. et al. (2005) Science 307, 1098-101.
  14. Jacinto, E. et al. (2006) Cell 127, 125-37.
Entrez-Gene Id
207 , 208 , 10000 , 66824 , 3059 , 6773
Swiss-Prot Acc.
P31749 , P31751 , Q9Y243 , Q9EPB4 , P14317 , P52630
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.

Upstream / Downstream

pathwayImage

Explore pathways related to this product.