Revision 1

#93195Store at -20C

1 Kit

(8 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
Cathepsin B (D1C7Y) XP® Rabbit mAb 31718 20 µl 44, 27, 24 kDa Rabbit IgG
HIF-1α (D1S7W) XP® Rabbit mAb 36169 20 µl 120 kDa Rabbit IgG
Hydroxy-HIF-1α (Pro564) (D43B5) XP® Rabbit mAb 3434 20 µl 120 kDa Rabbit IgG
Galectin-3/LGALS3 (D4I2R) XP® Rabbit mAb 87985 20 µl 28 kDa Rabbit IgG
Axl (C89E7) Rabbit mAb 8661 20 µl 138 kDa Rabbit IgG
CD68 (D4B9C) XP® Rabbit mAb 76437 20 µl Rabbit IgG
CD68 MultiMab® Rabbit mAb mix 86985 20 µl 70-80, 130-140 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 Neurodegeneration Module Antibody Sampler Kit provides an economical means of detecting proteins identified as markers of microglial activity during neurodegeneration 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).
CD68 is a common marker for macrophage lineage cells; with expression found in the lysosome making it a useful marker for activated phagocytic microglia (5). Galectin-3 has been shown to regulate inflammatory response in neurodegenerative diseases, released by microglia in response to inflammatory stimuli (6). Cathepsin B is a widely expressed cysteine peptidase located in the lysosome as well as processed and secreted, playing a role in microglial-mediated neuronal death (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). Axl is a receptor tyrosine kinase that binds Gas6, stimulating regulatory effects on microglial phagocytic response to inflammatory stimuli (9).

  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. Hopperton, K.E. et al. (2018) Mol Psychiatry 23, 177-98.
  6. Yip, P.K. et al. (2017) Sci Rep 7, 41689.
  7. Gan, L. et al. (2004) J Biol Chem 279, 5565-72.
  8. Zhang, Z. et al. (2011) Curr Med Chem 18, 4335-43.
  9. Grommes, C. et al. (2008) J Neuroimmune Pharmacol 3, 130-40.

Background References

    Trademarks and Patents

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

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    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.

    Revision 1
    #93195

    Microglia Neurodegeneration Module Antibody Sampler Kit

    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 1 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal cynomolgus monkey spleen using CD68 (D4B9C) XP® Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 2 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 Neurodegeneration Module Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from various cell lines using Cathepsin B (D1C7Y) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 4 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 Neurodegeneration Module Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from Hep G2 cells untreated (-) or treated with cobalt chloride (100 µM, 4 h; +), Raji cells untreated (-) or treated with cobalt chloride (100 µM, 4 h; +) and U-2 OS cells untreated (-) or treated with DMOG (1 mM, 6 h; +) using HIF-1α (D1S7W) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Microglia Neurodegeneration 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 Neurodegeneration Module Antibody Sampler Kit: Image 7 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 Neurodegeneration Module Antibody Sampler Kit: Image 8 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 Neurodegeneration Module Antibody Sampler Kit: Image 9 Expand Image
    Immunohistochemical analysis of paraffin-embedded human tonsil using CD68 (D4B9C) XP® Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 10 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 Neurodegeneration Module Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from THP-1, U-937, and Jurkat cells using CD68 MultiMab® Rabbit mAb mix (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from various cell lines using Galectin-3/LGALS3 (D4I2R) XP® Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing full-length human Cathepsin B (hCTSB; +) or mouse Cathespin B (mCTSB; +) using Cathepsin B (D1C7Y) XP® Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 14 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 Neurodegeneration Module Antibody Sampler Kit: Image 15 Expand Image
    Immunoprecipitation of HIF-1α from lysate of Hep G2 cells treated with cobalt chloride (100 µM, 4 h). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is HIF-1α (D1S7W) XP® Rabbit mAb. Western blot analysis was performed using HIF-1α (D1S7W) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 16 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse spleen using HS1 (D5A9) XP® Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 17 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 Neurodegeneration Module Antibody Sampler Kit: Image 18 Expand Image
    Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using CD68 (D4B9C) XP® Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 19 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 Neurodegeneration Module Antibody Sampler Kit: Image 20 Expand Image
    Immunoprecipitation of CD68 from U-937 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is CD68 MultiMab® Rabbit mAb mix. Western blot analysis was performed using CD68 MultiMab® Rabbit mAb mix.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 21 Expand Image
    Immunoprecipitation of Galectin-3/LGALS3 from SK-MEL-28 cell extracts. Lane 1 is 10% input, lane 2 is Normal Rabbit IgG #2729, and lane 3 is Galectin-3/LGALS3 (D4I2R) XP® Rabbit mAb (lane 3).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Cathepsin B (D1C7Y) XP(R) Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 23 Expand Image
    Confocal immunofluorescent analysis of Hep G2 cells, untreated (left) or treated with cobalt chloride (500 μM, 24 h; right), using HIF-1α (D1S7W) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded LL2 syngeneic tumor using HS1 (D5A9) XP® Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 25 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 Neurodegeneration Module Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using CD68 (D4B9C) XP® Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded breast carcinoma using Axl (C89E7) Rabbit mAb. Note staining of infiltrating cells.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Galectin-3/LGALS3 (D4I2R) XP® Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human kidney using Cathepsin B (D1C7Y) XP(R) Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 30 Expand Image
    Flow cytometric analysis of U-2 OS cells, untreated (blue) or treated with DMOG (1 mM, 6 h; green), using HIF-1α (D1S7W) 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 Neurodegeneration Module Antibody Sampler Kit: Image 31 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 Neurodegeneration Module Antibody Sampler Kit: Image 32 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse forestomach using ASC/TMS1 (D2W8U) Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 33 Expand Image
    Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using Axl (C89E7) Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded SK-MEL-28 (left) and LNCaP (right) cell pellets using Galectin-3/LGALS3 (D4I2R) XP® Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 35 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Cathepsin B (D1C7Y) XP(R) Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 36 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with cobalt chloride (100 μM) overnight and HIF-1α (D1S7W) XP® 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 ARRDC3, a known target gene of HIF-1α (see additional figure containing ChIP-qPCR data).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 37 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 Neurodegeneration Module Antibody Sampler Kit: Image 38 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using ASC/TMS1 (D2W8U) Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 39 Expand Image
    Multiplex immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using CD68 (D4B9C) XP® rabbit mAb (red), PD-1 (EH33) mouse mAb #43248 (green), CD8α (C8/144B) mouse mAb #70306 (magenta), Pan-keratin (C11) mouse mAb #4545 (cyan), LAG3 (D2G4O) XP® rabbit mAb #15372 (orange), and TIM-3 (D5D5R) XP® rabbit mAb #45208 (yellow).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 40 Expand Image
    Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using Axl (C89E7) Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 41 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Galectin-3/LGALS3 (D4I2R) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 42 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with cobalt chloride (100 μM) overnight and HIF-1α (D1S7W) XP® 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 5 (upper), including ARRDC3 (lower), a known target gene of HIF-1α (see additional figure containing ChIP-qPCR data).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 43 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 Neurodegeneration Module Antibody Sampler Kit: Image 44 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 Neurodegeneration Module Antibody Sampler Kit: Image 45 Expand Image
    Multiplex immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using CD68 (D4B9C) XP® rabbit mAb (orange), PD-L1 (E1L3N®) XP® rabbit mAb #13684 (red), PD-L2 (D7U8C) XP® rabbit mAb (magenta) #82723, Arginase-1 (D4E3M) XP® rabbit mAb #93668 (green), IDO (D5J4E) rabbit mAb #86630 (yellow), and Pan-keratin (C11) mouse mAb #4545 (cyan).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 46 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using Axl (C89E7) Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 47 Expand Image
    Confocal immunofluorescent analysis of SK-MEL-28 (left) and LNCaP (right) cells, using Galectin-3/LGALS3 (D4I2R) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 48 Expand Image
    Confocal immunofluorescent analysis of Malme-3M (left) and MCF7 (right) cells using Cathepsin (D1C7Y) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 49 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with cobalt chloride (100 μM, overnight) and either HIF-1α (D1S7W) XP® 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 ARRDC3 downstream primers, SimpleChIP® Human ERRFI1 Upstream Primers #31180, 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 Neurodegeneration Module Antibody Sampler Kit: Image 50 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 Neurodegeneration Module Antibody Sampler Kit: Image 51 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse thymus using ASC/TMS1 (D2W8U) Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 52 Expand Image
    Immunohistochemical analysis of paraffin-embedded THP-1 (left) and Jurkat (right) cell pellets using CD68 (D4B9C) XP® Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 53 Expand Image
    Immunohistochemical analysis of paraffin-embedded metastatic lung carcinoma using Axl (C89E7) Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 54 Expand Image
    Flow cytometric analysis of fixed/permeabilized Daudi cells (blue, negative) and SK-MEL-28 cells (green, positive) using Cathepsin B (D1C7Y) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 55 Expand Image
    CUT&RUN was performed with MCF7 cells treated with cobalt chloride (100 μM) overnight and HIF-1α (D1S7W) XP® 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 STC2, a known target gene of HIF-1α (see additional figure containing CUT&RUN-qPCR data).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 56 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse small intestine using ASC/TMS1 (D2W8U) Rabbit mAb.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 57 Expand Image
    Confocal immunofluorescent analysis of THP-1 (left) and Jurkat (right) cells using CD68 (D4B9C) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Projected images from a z-stack series are shown.
    Microglia Neurodegeneration 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 Neurodegeneration Module Antibody Sampler Kit: Image 59 Expand Image
    CUT&RUN was performed with MCF7 cells treated with cobalt chloride (100 μM) overnight and HIF-1α (D1S7W) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 5 (upper), including STC2 (lower), a known target gene of HIF-1α (see additional figure containing CUT&RUN-qPCR data).
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 60 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 Neurodegeneration Module Antibody Sampler Kit: Image 61 Expand Image
    Flow cytometric analysis of live human peripheral blood mononuclear cells co-stained with anti-human CD14 using CD68 (D4B9C) XP® Rabbit mAb (right) compared to a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 62 Expand Image
    CUT&RUN was performed with MCF7 cells treated with cobalt chloride (100 μM) overnight and HIF-1α (D1S7W) XP® 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 STC2 exon 1 primers, human FAM162A 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.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 63 Expand Image
    Flow cytometric analysis of fixed and permeabilized human peripheral blood mononuclear cells co-stained with anti-human CD14 using CD68 (D4B9C) XP® Rabbit mAb (right) compared to a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 64 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 Neurodegeneration Module Antibody Sampler Kit: Image 65 Expand Image
    Flow cytometric analysis of fixed and permeabilized Jurkat cells (blue, negative) and THP-1 cells (green, positive) using CD68 (D4B9C) 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.
    Microglia Neurodegeneration Module Antibody Sampler Kit: Image 66 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 Neurodegeneration Module Antibody Sampler Kit: Image 67 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 Neurodegeneration Module Antibody Sampler Kit: Image 68 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 Neurodegeneration Module Antibody Sampler Kit: Image 69 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.