The Microglia Proliferation Module Antibody Sampler Kit provides an economical means of detecting proteins identified as markers of microglial proliferation by western blot and/or immunofluorescence.
Specificity / Sensitivity
Each antibody in the Microglia Proliferation 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. Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11) XP® Rabbit mAb detects endogenous levels of Aurora A/B/C only when phosphorylated at either Thr288, Thr232, or Thr198 respectively. HP1α/β (C7F11) Rabbit mAb does not cross-react with HP1 γ proteins.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu310 of mouse HS1, Cys60 of human Survivin, Thr232 of human Aurora B, the carboxy terminus of human HP1α, the amino terminus of human Ki-67 and MCM2, and recombinant mouse ASC/TMSI 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).
Ki-67 is a nuclear nonhistone protein (5) universally expressed among proliferating cells and absent in quiescent cells (6). Minichromosome maintenance protein 2 (MCM2) is a nuclear protein that plays a role in DNA replication and cell division (7) and is commonly used as a marker for cell proliferation, including brain tissue (8). Survivin binds and inhibits caspase-3, controlling the checkpoint in the G2/M-phase of the cell cycle by inhibiting apoptosis and promoting cell division (9). Aurora A, B, and C are a family of highly conserved serine/threonine kinases that regulate chromosomal alignment and segregation during mitosis and meiosis. Their activity requires autophosphorylation of a threonine within their kinase domain at site Thr288 of Aurora A, Thr232 of Aurora B, and Thr198 of Aurora C (10). Heterochromatin protein 1 (HP1) α and β are heterochromatic adaptor molecules involved in both gene silencing and higher order chromatin structure (11).