Revision 1

#83163Store 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
ASC/TMS1 (D2W8U) Rabbit mAb 67824 20 µl 22 kDa Rabbit IgG
HS1 (D5A9) XP® Rabbit mAb 3892 20 µl 80 kDa Rabbit IgG
Ki-67 (D3B5) Rabbit mAb 9129 20 µl Rabbit IgG
Axl (C89E7) Rabbit mAb 8661 20 µl 138 kDa Rabbit IgG
Hydroxy-HIF-1α (Pro564) (D43B5) XP® Rabbit mAb 3434 20 µl 120 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
Lamin A/C (4C11) Mouse mAb 4777 20 µl 74 (Lamin A), 63 (Lamin C) kDa Mouse IgG2a
IQGAP1 (D8K4X) XP® Rabbit mAb 20648 20 µl 195 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 Microglia Cross Module Antibody Sampler Kit provides an economical means of detecting proteins identified as markers of microglial activity corresponding to proliferation, neurodegeneration, interferon and LPS-relation by western blot and/or immunofluorescence.

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 antibody.

Background

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).
Ki-67 is a nuclear nonhistone protein (5) universally expressed among proliferating cells and absent in quiescent cells (6). Axl is a receptor tyrosine kinase that binds Gas6, stimulating regulatory effects on microglial phagocytic response to inflammatory stimuli (7). Hypoxia inducible factor-1 (HIF-1α) is a transcription factor responsible for adaptation to low oxygen environments whose downstream effects have been shown in a number of neurodegenerative diseases. Under normoxic conditions, HIF-1α is proline hydroxylated leading to ubiquitin mediated degradation (8). Stat2 is critical to the transcriptional responses induced by type I interferons, IFN-alpha/beta (9,10). In response to IFN-alpha/beta, Stat2 is activated by phosphorylation at site Tyr690 through associations with receptor-bound Jak kinases (11). Lamins are nuclear membrane structural components important for maintaining normal cell functions. Lamin A/C is cleaved by caspase-6 and serves as a marker for caspase-6 activation. The cleavage of lamins results in nuclear dysregulation and cell death (12,13). IQGAP1 is ubiquitously expressed and has been found to interact with APC (14) and the CLIP170 complex in response to small GTPases, promoting cell polarization and migration (15).

  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. Gerdes, J. et al. (1983) Int J Cancer 31, 13-20.
  6. Weigel, M.T. and Dowsett, M. (2010) Endocr Relat Cancer 17, R245-62.
  7. Grommes, C. et al. (2008) J Neuroimmune Pharmacol 3, 130-40.
  8. Zhang, Z. et al. (2011) Curr Med Chem 18, 4335-43.
  9. Fu, X.Y. et al. (1992) Proc Natl Acad Sci U S A 89, 7840-3.
  10. Ihle, J.N. (2001) Curr Opin Cell Biol 13, 211-7.
  11. Improta, T. et al. (1994) Proc Natl Acad Sci U S A 91, 4776-80.
  12. Oberhammer, F.A. et al. (1994) J Cell Biol 126, 827-37.
  13. Rao, L. et al. (1996) J Cell Biol 135, 1441-55.
  14. Watanabe, T. et al. (2004) Dev Cell 7, 871-83.
  15. Fukata, M. et al. (2002) Cell 109, 873-85.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

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    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
    #83163

    Microglia Cross Module Antibody Sampler Kit

    Microglia Cross Module Antibody Sampler Kit: Image 1 Expand Image
    Flow cytometric analysis of Daudi cells (blue) and MJ cells (green) using Lamin A/C (4C11) Mouse mAb (solid lines) or concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
    Microglia Cross Module Antibody Sampler Kit: Image 2 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.
    Microglia Cross Module Antibody Sampler Kit: Image 3 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells using Axl (C89E7) Rabbit mAb #8661. The virtual lane view (left) shows a single 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.
    Microglia Cross Module Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from various cell lines using IQGAP1 (D8K4X) XP® Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from HeLa cells, treated with either 10 μM of MG132 (to accumulate hydroxylated HIF-1α) or 10 µM MG132 and 1 mM DMOG (to accumulate nonhyroxylated HIF-1α), using Hydroxy-HIF-1α (Pro564) (D43B5) XP® Rabbit mAb (upper) or total HIF-1α Antibody #3716 (lower).
    Microglia Cross Module Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of cell extracts from Baf3, 32D, and mouse spleen using HS1 (D5A9) XP® Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from control HeLa cells (lane 1) or HeLa cells with an apparent in-frame truncation mutation in the gene encoding LMNA (lane 2) using Lamin A/C (4C11) Mouse mAb #4777 (upper) or α-actinin (D6F6) XP® Rabbit mAb #6487 (lower). The change in LMNA molecular weight in the mutated HeLa cells is consistent with an in-frame deletion.
    Microglia Cross Module Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from J774A.1 and Raw 264.7 cells using ASC/TMS1 (D2W8U) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Microglia Cross Module Antibody Sampler Kit: Image 9 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.
    Microglia Cross Module Antibody Sampler Kit: Image 10 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.
    Microglia Cross Module Antibody Sampler Kit: Image 11 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.
    Microglia Cross Module Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of HeLa Cell Extracts, untreated (-) or Axl knock-out (+) using Axl (C89E7) Rabbit mAb, #8661 (upper) or GAPDH (D16H11) XP® Rabbit mAb, #5174 (lower).
    Microglia Cross Module Antibody Sampler Kit: Image 13 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).
    Microglia Cross Module Antibody Sampler Kit: Image 14 Expand Image
    Confocal immunofluorescent analysis of the ventricular zone in P21 mouse brain using Ki-67 (D3B5) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Microglia Cross Module Antibody Sampler Kit: Image 15 Expand Image
    breast:

    Immunohistochemical analysis of paraffin-embedded human infiltrating papillary carcinoma of the breast using IQGAP1 (D8K4X) XP® Rabbit mAb.

    Microglia Cross Module Antibody Sampler Kit: Image 16 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, treated with either 10 μM MG132 (left) or 10 μM MG132 and 1 mM DMOG (right), using Hydroxy-HIF-1α (Pro564) (D43B5) XP® Rabbit mAb (green). Actin filaments have been labeled using DY-554 phalloidin (red).
    Microglia Cross Module Antibody Sampler Kit: Image 17 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse spleen using HS1 (D5A9) XP® Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 18 Expand Image
    Western blot analysis of extracts from various cell lines using Lamin A/C (4C11) Mouse mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 19 Expand Image
    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. Western blot analysis was performed using ASC/TMS1 (D2W8U) Rabbit mAb.
    Microglia Cross Module 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.
    Microglia Cross Module 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).
    Microglia Cross Module Antibody Sampler Kit: Image 22 Expand Image
    Western blot analysis of extracts from various cell lines using Axl (C89E7) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Microglia Cross Module Antibody Sampler Kit: Image 23 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).
    Microglia Cross Module Antibody Sampler Kit: Image 24 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Ki-67 (D3B5) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Microglia Cross Module Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded SK-MEL-28 (left) and LNCaP (right) cell pellets using IQGAP1 (D8K4X) XP® Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded LL2 syngeneic tumor using HS1 (D5A9) XP® Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 27 Expand Image
    Western blot analysis of extracts from THP-1 cells, untreated or treated with cycloheximide (CHX, 10 μg/ml, overnight) followed by TNF-α #8902 (20 ng/ml, 4 hr), using Lamin A/C (4C11) Mouse mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded J774A.1 cell pellet (left, positive) or RAW 264.7 cell pellet (right, negative) using ASC/TMS1 (D2W8U) Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 29 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).
    Microglia Cross Module Antibody Sampler Kit: Image 30 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.
    Microglia Cross Module Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded breast carcinoma using Axl (C89E7) Rabbit mAb. Note staining of infiltrating cells.
    Microglia Cross Module 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.
    Microglia Cross Module Antibody Sampler Kit: Image 33 Expand Image
    Flow cytometric analysis of Jurkat cells using Ki-67 (D3B5) Rabbit mAb and Propidium Iodide (PI)/RNase Staining Solution #4087. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Microglia Cross Module Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using IQGAP1 (D8K4X) XP® Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 35 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse cortex from wild-type (left) or an amyloid mouse model of Alzheimer's disease (right) using HS1 (D5A9) XP® Rabbit mAb (green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using β-Amyloid (D54D2) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) #35363 (red) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Microglia Cross Module Antibody Sampler Kit: Image 36 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Lamin A/C (4C11) Mouse mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 37 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse forestomach using ASC/TMS1 (D2W8U) Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 38 Expand Image
    Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using Axl (C89E7) Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 39 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using IQGAP1 (D8K4X) XP® Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 40 Expand Image
    Confocal immunofluorescent analysis of mouse Tg2576 brain which overexpresses mutant human APP695. Sections were first labeled with HS1 (D5A9) XP® Rabbit mAb (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).
    Microglia Cross Module Antibody Sampler Kit: Image 41 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Lamin A/C (4C11) Mouse mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 42 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using ASC/TMS1 (D2W8U) Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 43 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.
    Microglia Cross Module Antibody Sampler Kit: Image 44 Expand Image
    Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using Axl (C89E7) Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 45 Expand Image
    Confocal immunofluorescent analysis of A549 (left) and Hep G2 (right) cells using IQGAP1 (D8K4X) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Microglia Cross Module Antibody Sampler Kit: Image 46 Expand Image
    Confocal immunofluorescent analysis of 32D cells (left) and C2C12 cells (right), using HS1 (D5A9) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Microglia Cross Module Antibody Sampler Kit: Image 47 Expand Image
    Immunofluorescent analysis of normal rat brain using Lamin A/C (4C11) Mouse mAb (green) and MAP2 Antibody #4542 (red).
    Microglia Cross Module Antibody Sampler Kit: Image 48 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse colon using ASC/TMS1 (D2W8U) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    Microglia Cross Module Antibody Sampler Kit: Image 49 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).
    Microglia Cross Module Antibody Sampler Kit: Image 50 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using Axl (C89E7) Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 51 Expand Image
    Flow cytometric analysis of NIH/3T3 cells (blue, negative) and 32D clone 3 cells (green, positive) using HS1 (D5A9) XP® Rabbit mAb (solid lines) or 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.
    Microglia Cross Module Antibody Sampler Kit: Image 52 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Lamin A/C (4C11) Mouse mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
    Microglia Cross Module Antibody Sampler Kit: Image 53 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse thymus using ASC/TMS1 (D2W8U) Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 54 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.
    Microglia Cross Module Antibody Sampler Kit: Image 55 Expand Image
    Immunohistochemical analysis of paraffin-embedded metastatic lung carcinoma using Axl (C89E7) Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 56 Expand Image
    Flow cytometric analysis of HeLa cells (green) using Lamin A/C (4C11) Mouse mAb (solid lines) or a concentration matched Mouse (G3A1) mAb IgG Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
    Microglia Cross Module Antibody Sampler Kit: Image 57 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse small intestine using ASC/TMS1 (D2W8U) Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 58 Expand Image
    Immunohistochemical analysis of paraffin-embedded cell pellets, NCI-H1299 (left) or Jurkat (right), using Axl (C89E7) Rabbit mAb.
    Microglia Cross Module Antibody Sampler Kit: Image 59 Expand Image
    Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor (top left), 4T1 syngeneic mammary tumor (top right), Renca cell pellet (bottom left), and 4T1 cell pellet (bottom right) using ASC/TMS1 (D2W8U) Rabbit mAb. Both tumors show staining of infiltrating immune cells. Note the presence of staining in the Renca tumor cells and the lack of staining in the 4T1 tumor cells consistent with staining results on corresponding cell pellets.
    Microglia Cross Module Antibody Sampler Kit: Image 60 Expand Image
    Confocal immunofluorescent analysis of DU 145 (left) and HCC827 (right) cells using Axl (C89E7) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Microglia Cross Module Antibody Sampler Kit: Image 61 Expand Image
    Flow cytometric analysis of fixed and permeabilized Jurkat cells (blue, negative) and DU145 cells (green, positive) using Axl (C89E7H4) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Microglia Cross Module Antibody Sampler Kit: Image 62 Expand Image
    Confocal immunofluorescent analysis of mouse Tg2576 brain which overexpresses mutant human APP695. Sections were first labeled with ASC/TMS1 (D2W8U) Rabbit mAb #67824 (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).
    Microglia Cross Module Antibody Sampler Kit: Image 63 Expand Image
    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 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Note the translocation of ASC to inflammasomes following stimulation with LPS and ATP (white arrows).
    Microglia Cross Module Antibody Sampler Kit: Image 64 Expand Image
    Flow cytometric analysis of Raw264.7 cells (blue) and J774A.1 cells (green) using ASC/TMS1 (D2W8U) Rabbit mAb (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.
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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.