Microglia Neurodegeneration Module Antibody Sampler Kit #93195
Product Information
Kit Usage Information
Protocols
- 3434: Western Blotting, Immunoprecipitation (Magnetic), Immunofluorescence
- 3892: Western Blotting, Immunoprecipitation (Magnetic), Immunohistochemistry (Paraffin), Immunofluorescence, Immunofluorescence, Flow
- 7074: Western Blotting
- 8661: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
- 31718: Western Blotting, Immunohistochemistry (Paraffin), Immunofluorescence*, Flow
- 36169: Western Blotting, Immunoprecipitation (Magnetic), Immunofluorescence, Flow, ChIP Magnetic, Chromatin IP-seq, CUT&RUN Assay
- 67824: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Immunofluorescence, Flow
- 76437: Immunohistochemistry (Paraffin), Immunofluorescence*, Flow, Flow Cytometry Live Cell Unconjugated Rabbit
- 86985: Western Blotting, Immunoprecipitation (Agarose)
- 87985: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence
Product Description
Specificity / Sensitivity
Each antibody in the Microglia Neurodegeneration Module Antibody Sampler Kit detects endogenous levels of its target protein. 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. Cathepsin B (D1C7Y) XP® Rabbit mAb recognizes endogenous levels of total cathepsin B protein and detects the heavy chain subunit of cathepsin B. Hydroxy-HIF-1α (Pro564) (D43B5) XP® Rabbit mAb detects endogenous levels of HIF-1α only when hydroxylated at Pro564 and may cross-react with other overexpressed proline hydroxylated proteins. Axl (C89E7) Rabbit mAb does not cross-react with Tyro3.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu310 of mouse HS1, Leu478 of human HIF-1α, the amino terminus of human Galectin-3/LGALS3, a hydroxypeptide surrounding Pro564 of human HIF-1α, and recombinant proteins specific to mouse ASC/TMS1, human Axl, human CD68, and the heavy chain subunit of human cathepsin B protein.
MultiMab™ rabbit monoclonal mix antibodies are prepared by combining individual rabbit monoclonal clones in optimized ratios for the approved applications. This product is optimized to detect CD68 as a monomer and a dimer by western blot and was produced by immunizing animals with recombinant human CD68 protein.
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).
- Friedman, B.A. et al. (2018) Cell Rep 22, 832-47.
- Zhang, Y. et al. (2014) J Neurosci 34, 11929-47.
- Kitamura, D. et al. (1995) Biochem Biophys Res Commun 208, 1137-46.
- Srinivasula, S.M. et al. (2002) J Biol Chem 277, 21119-22.
- Hopperton, K.E. et al. (2018) Mol Psychiatry 23, 177-98.
- Yip, P.K. et al. (2017) Sci Rep 7, 41689.
- Gan, L. et al. (2004) J Biol Chem 279, 5565-72.
- Zhang, Z. et al. (2011) Curr Med Chem 18, 4335-43.
- Grommes, C. et al. (2008) J Neuroimmune Pharmacol 3, 130-40.
Limited Uses
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