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Commit: f2d32940205a64f990b886d724ccee2c9935daff
XML generation date: 2024-11-20 18:01:13.529
Product last modified at: 2024-09-20T07:01:40.763Z
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PDP - Template Name: Antibody Sampler Kit
PDP - Template ID: *******4a3ef3a

Microglia Cross Module Antibody Sampler Kit #83163

    Product Information

    Product 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.

    Specificity / Sensitivity

    Each antibody in the Microglia Cross Module Antibody Sampler Kit detects endogenous levels of its target protein. Hydroxy-HIF-1α (Pro564) (D43B5) XP® Rabbit mAb detects endogenous levels of HIF-1α only when hydroxylated at Pro564. This antibody may cross-react with other overexpressed proline hydroxylated proteins. Phospho-Stat2 (Tyr690) (D3P2P) Rabbit mAb recognizes endogenous levels of Stat2 protein only when phosphorylated at Tyr690. Axl (C89E7) Rabbit mAb does not cross-react with Tyro3. 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. Lamin A/C (4C11) Mouse mAb detects endogenous levels of lamin A and lamin C proteins. It also reacts with the larger fragments of lamin A (50 kDa) and lamin C (41 kDa) produced by caspase cleavage during apoptosis. This antibody does not cross-react with lamins B1 and B2.

    Source / Purification

    Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu310 of mouse HS1, Pro564 of human HIF-1α, Leu706 of human Stat2, Tyr690 of human Stat2, the amino terminus of human Ki-67 and IQGAP1, and recombinant fragment of human Axl, human lamin A, and mouse ASC/TMS1.

    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.
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