Apoptosis and autophagy |
Caspase-3 |
Caspase-3 is an intracellular proteases that mediates cell death and plays a critical role in apoptosis. Activation of caspase-3 requires proteolytic processing (Fernandes-Alnemri et al., 1994). |
Sigma–Aldrich, anti-Caspase 3, #C8487-200UL |
1:1000 |
|
LC3 |
Light chain 3 (LC3) is an autophagy marker. Cleavage of LC3 yields the cytosolic LC3-I form. During autophagy, LC3-I is converted to LC3-II permitting LC3 to become associated with autophagic vesicles (Kabeya et al., 2000). |
Sigma–Aldrich, anti-LC3, #L8918-200UL |
1:1000 |
Neuronal |
NeuN |
Neuronal nuclei (NeuN) is a nuclear protein expressed in most post-mitotic neurons of the central and peripheral nervous systems (Mullen et al., 1992). Marker for mature neurons. |
Cell Signaling, anti-NeuN (D3S31), #12943 |
1:2000 |
|
Synaptophysin |
Synaptophysin is an integral membrane protein of small synaptic vesicles in the brain (Wiedenmann et al., 1986). |
Millipore, anti-Synaptophysin, #MAB329 |
1:10,000 |
|
PSD95 |
Postsynaptic density protein 95 (PSD95) is a scaffolding protein involved in the assembly and function of the postsynaptic density complex (Chetkovich et al., 2002). |
Cell Signaling, anti-Postsynaptic density protein 95, #2507 |
1:1000 |
Glial |
GFAP |
Glial fibrillary acidic protein (GFAP) forms intermediate filaments in astroglial cells and modulate their motility and shape. GFAP filaments are characteristic of differentiated and mature brain astrocytes (Eng et al., 2000). |
Cell Signaling, anti-Glial Fibrillary Acidic Protein (GA5), #3670 |
1:5000 |
|
ALDH1L1 |
10-Formyltetrahydrofolate dehydrogenase (ALDH1L1) is a multidomain protein that serves as a CNS astrocyte marker (Cahoy et al., 2008). |
Millipore, anti-ALDH1L1 (N103/39), #MABN495 |
1:2000 |
|
CD11b |
Cluster of differentiation molecule 11b (CD11b) is a transmembrane protein expressed by neutrophils, monocytes, macrophages, and microglia (Solovjov et al., 2005). |
Abcam, anti-CD11b, #ab75476 |
1:2000 |
Vascular |
VEGFA |
Growth factor active in angiogenesis, vasculogenesis, and endothelial cell growth. Induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis, and induces permeabilization of blood vessels (Leung et al., 1989). |
Abcam, anti-VEGFA #ab46154 |
1:1000 |
|
CD31 (PECAM-1) |
CD31 (Platelet Endothelial Cell Adhesion Molecule-1, PECAM-1) is a cell adhesion molecules expressed by circulating platelets, monocytes, neutrophils, some T cells, and makes up a large portion of endothelial cell intercellular junctions. Modulates cell adhesion, endothelial cell migration, and angiogenesis (Newman, 1997). |
Cell Signaling, anti-CD31 (PECAM-1) (D8V9E) #77699 Sigma–Aldrich, anti-PECAM-1 #SAB4502167 |
1:1000 1:1000 |
|
Collagen IV |
Type IV collagen is the major structural component of basement membranes. Type IV collagen is a network-forming collagen that provides a molecular scaffold and interacts with cells, growth factors, and other basement membrane components such as laminin, nidogen, and perlecan (Kuhn, 1995). |
Abcam, anti-Collagen IV #ab6586 |
1:1000 |
Neurodegeneration |
Amyloid-β |
Amyloid-β (Aβ) peptide produced through sequential proteolytic processing of amyloid precursor protein, and it is prone to aggregate in pathological conditions (Haass and Selkoe, 2007; Walsh and Selkoe, 2007). |
Biolegend, anti-Amyloid-β (6E10), # SIG-39320 |
1:1000 |
|
α-Synuclein |
α-Synuclein (α-Syn) is expressed in brain, primarily in presynaptic nerve terminals. Although the exact function has not been determined, it has been linked to the prominent neurodegenerative disorders (Maroteaux et al., 1988; van der Putten et al., 2000). |
BD Bioscience, anti-α-Synuclein, #610787 |
1:1000 |
|
Tau5 |
Tau5 detects total levels of Tau. Tau are a microtubule-associated proteins that bind to the tubulin subunits of microtubule structures, and promote and stabilize microtubule assembly (Avila et al., 2004; Johnson and Stoothoff, 2004). |
Millipore, anti-Tau5, #MAB361 |
1:2000 |
|
P-Tau (Ser400/ Thr403/Ser404) |
Phospho-Tau (P-Tau) recognizes endogenous levels of tau protein when phosphorylated at Ser400 or Thr403 or Ser404. Studies have shown that tau phosphorylation at Ser404 destabilizes microtubules and that tau is hyperphosphorylated at Ser404 in Alzheimer’s disease (Evans et al., 2000). Phosphorylation decreases the ability of tau to bind to microtubules, destabilizing the structure and driving it toward disassembly. Neurofibrillary tangles are a major hallmark of Alzheimer’s disease; these tangles are composed of hyperphosphorylated tau (Johnson and Stoothoff, 2004). |
Cell Signaling, anti-Phospo-Tau (Ser400/Thr403/Ser404), #11837 |
1:1000 |
|
P-Tau (Ser396) |
Phospho-Tau (Ser396) (P-Tau Ser396) detects endogenous levels of Tau only when phosphorylated at serine 396. Phosphorylation at Ser396 has shown to destabilize microtubules and contribute to different neurological disorders (Bramblett et al., 1993; Evans et al., 2000). |
Cell Signaling, anti-Phospo-Tau (Ser396), #9632 |
1:1000 |
Housekeeping |
β-Actin |
β-Actin is a cytoskeletal housekeeping protein. |
Sigma–Aldrich, monoclonal anti-β-actin-HRP antibody, A3854 |
1:50,000 |
Secondary |
Rabbit IgG |
Secondary antibody. |
Bio-Rad, Anti-Rabbit-HRP antibody, #170-6515 |
1:7500 |
|
Mouse IgG |
Secondary antibody. |
Bio-Rad, Anti-Mouse-HRP antibody, #170-6516 |
1:10,000 |