The Mouse Reactive Alzheimer's Disease Model Microglia Phenotyping IF Antibody Sampler Kit provides an economical means of detecting microglia proteins in β-Amyloid mouse models of Alzheimer’s Disease (AD) 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 Reactive Alzheimer's Disease Model Microglia Phenotyping IF Antibody Sampler Kit detects endogenous levels of its target protein. β-Amyloid (D54D2) XP® Rabbit mAb detects transgenically expressed human APP in mouse models and several isoforms of Aβ, such as Aβ-37, Aβ-38, Aβ-39, Aβ-40, and Aβ-42. 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 detects the heavy chain subunit of cathepsin B. Cathepsin D (E179) Antibody detects endogenous levels of preprocathepsin D, procathepsin D, and the heavy chain subunit of mature cathepsin D protein.
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, Ala1153 of mouse CD11c protein, and Glu179 of mouse cathepsin D protein. Antibodies are also produced with the recombinant heavy chain subunit of human cathepsin B protein and mouse ASC/TMS1 protein, and the amino terminus of human β-amyloid peptide (Aβ), human TMEM119, and mouse GPNMB protein. Cathepsin D (E179) Antibody is purified by peptide affinity chromatography.
Distinct microglial activation states have been identified using RNA-seq data from a vast array of neurological disease and aging models. In both mouse models of Alzheimer’s Disease (AD) and AD patients, unique microglia molecular signatures are associated with disease progression (1-3). AD progression is correlated with the extracellular deposition and accumulation of the released Aβ fragments, derived from the transmembrane glycoprotein Amyloid β (Aβ) precursor protein (APP), that form amyloid plaques, the pathological hallmark of AD (4). Microglia are the resident macrophages of the brain and contribute to neurodegenerative disease (5). 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 (6,7). HS1 (HCLS1, LckBP1, p75) is a protein kinase substrate that is expressed only in tissues and cells of hematopoietic origin and is also expressed in microglia (8,9). Transmembrane protein 119 (TMEM119) is a cell-surface protein of unknown function, expressed exclusively by the microglia subset of myeloid and neural cells (10). Iba1+ microglia with both ramified and amoeboid morphologies express TMEM119, while Iba1+ macrophages are TMEM119 negative (11). TMEM119 and other homeostatic genes have been shown to be downregulated in microglia. In addition to general markers of microglia, several microglia genes are upregulated during disease progression (12). CD11c (integrin αX, ITGAX) is a transmembrane glycoprotein that forms an α/β heterodimer with CD18 (integrin β2), which interacts with a variety of extracellular matrix molecules and cell surface proteins (13). CD11c-positive microglia transcriptionally correlate with amyloid plaques (14). In addition, other genes are upregulated in a similar manner. Glycoprotein non-metastatic gene B (GPNMB) is a type I transmembrane glycoprotein overexpressed in many types of cancer. The GPNMB glycoprotein is involved in many physiological processes, including mediating transport of late melanosomes to keratinocytes (9,15). Cathepsin B and D are widely expressed cysteine and aspartyl proteases, respectively, involved in the normal degradation of proteins (16,17). 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 and may directly contribute to amyloid plaque formation (18,19).