The Mouse Microglia Marker IF Antibody Sampler Kit provides an economical means of detecting proteins identified as microglia markers by immunofluorescence and/or western blot. This kit includes enough primary antibodies to perform at least twenty IF-F tests or two western blot experiments per primary antibody.
Specificity / Sensitivity
Each antibody in the Mouse Microglia Marker IF Antibody Sampler Kit detects endogenous levels of its target protein. HS1 has a calculated size of 54 kDa, but has an apparent molecular weight of 80 kDa on SDS-PAGE gels. CD45 (30-F11) Rat mAb and CD11b/ITGAM (M1/70) Rat mAb detect an epitope within the extracellular domain. CD68 (E3O7V) Rabbit mAb has shown staining of uncertain specificity in mouse endometrial epithelium by immunohistochemistry.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Ala139 of human Iba1/AIF-1 protein, Leu310 of mouse HS1 protein, Gln260 of mouse F4/80 protein, the amino terminus of human TMEM119 and Ki-67 proteins, and recombinant mouse ASC/TMS1 protein and mouse CD68 that reacts with an epitope surrounding Gly113. CD11b/ITGAM (M1/70) Rat mAb and CD45 (30-F11) Rat mAb were purified from tissue culture supernatant via affinity chromatography.
Microglia are the resident macrophages of the central nervous system, responsible for immune response and maintenance of CNS homeostasis (1). Ionized calcium-binding adaptor molecule 1 (Iba1), also known as allograft inflammatory factor 1 (AIF-1), is uniquely expressed in cells of monocytic lineage and is, therefore, widely used as a marker for microglia/macrophages in the brain and other tissue (2,3). Transmembrane protein 119 (TMEM119) is a cell-surface protein of unknown function, expressed exclusively by the microglia subset of myeloid and neural cells (4). Iba1+ microglia with both ramified and amoeboid morphologies express TMEM119, while Iba1+ macrophages are TMEM119 negative (5). TMEM119 and other homeostatic genes have been shown to be downregulated in disease-associated microglia (DAM) (6). Cluster of differentiation molecule 11b (CD11b)/Integrin alpha M (ITGAM) is a transmembrane protein expressed by, and commonly used as a marker for, myeloid lineage cells, including neutrophils, monocytes, macrophages, and microglia (7). F4/80 (EMR1) is a heavily glycosylated G-protein-coupled receptor and is a well-established marker for mouse macrophages (8-10). Expression of F4/80 has been observed in microglia and subset populations of dendritic cells (11). The protein phosphatase (PTP) receptor CD45 is a type I transmembrane protein expressed in all nucleated hematopoietic cells (12). Studies suggest CD45 plays a role in regulation of microglial activation (13,14). CD68 (macrosialin) is a heavily glycosylated transmembrane protein that is expressed by and commonly used as a marker for monocytes and macrophages (15,16). It localizes to the lysosomal membrane and is upregulated during microglial activation (17,18). Ki-67 is a nuclear nonhistone protein (19) universally expressed among proliferating cells and absent in quiescent cells (20). 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 (21). HS1 is a protein kinase substrate that is expressed only in tissues and cells of hematopoietic origin (22) 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 (23).