|36422T||1 Kit (9 x 20 microliters)||
|ASC/TMS1 (D2W8U) Rabbit mAb 67824||20 µl||
|HS1 (D5A9) XP® Rabbit mAb 3892||20 µl||
||M R||80||Rabbit IgG|
|Rab11FIP1 (D9D8P) Rabbit mAb 12849||20 µl||
||H M Mk||85||Rabbit IgG|
|Integrin α4 (D2E1) XP® Rabbit mAb 8440||20 µl||
||H M R||70, 140, 150,||Rabbit IgG|
|IQGAP1 (D8K4X) XP® Rabbit mAb 20648||20 µl||
||H M R Mk||195||Rabbit IgG|
|Cleaved Lamin A (Small Subunit) (30H5) Mouse mAb 2036||20 µl||
||H M R||28||Mouse IgG1|
|IKKε (D61F9) XP® Rabbit mAb 3416||20 µl||
||M R||80||Rabbit IgG|
|Lamin A/C (4C11) Mouse mAb 4777||20 µl||
||H M R Mk||74 (Lamin A), 63 (Lamin C)||Mouse IgG2a|
|Phospho-Ezrin (Thr567)/Radixin (Thr564)/Moesin (Thr558) (48G2) Rabbit mAb 3726||20 µl||
||H M R Mk||75 Moesin. 80 Ezrin, Radixin.||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu310 of mouse HS1, Leu248 of human Rab11FIP1, Ser1027 of human integrin α4, Thr567 of human ezrin, Asp230 of human lamin A, the amino terminus of human IQGAP1, the carboxy terminus of mouse IKKε, and a recombinant fragment of human lamin A and mouse ASC/TMS1 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).
The Rab11-family interacting proteins (Rab11-FIPs) facilitate Rab11-dependent vesicle recycling through interaction with the conserved carboxyl terminal Rab11 binding domain (5,6). Rab11FIP1 has been shown to play a role in endocytic sorting and trafficking of EGFR and integrin subunits (6). Integrins are α/β heterodimeric cell surface receptors that mediate cell adhesion and migration and regulate cell growth and survival. Two significant α4 integrins, α4β1 and α4β7, interact with VCAM-1, fibronectin, and MAdCAM-1 at cell adhesions and have been shown to play an important role in cell trafficking during inflammatory processes (7-9). 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 (10,11). The ezrin, radixin, and moesin (ERM) proteins function as linkers between the plasma membrane and the actin cytoskeleton and are involved in cell adhesion, membrane ruffling, and microvilli formation (12). ERM proteins undergo intra or intermolecular interaction between their amino- and carboxy-terminal domains, existing as inactive cytosolic monomers or dimers (13). Phosphorylation at a carboxy-terminal threonine residue (Thr567 of ezrin, Thr564 of radixin, Thr558 of moesin) disrupts the amino- and carboxy-terminal association and may play a key role in regulating ERM protein conformation and function (14,15). IQGAPs are scaffolding proteins involved in mediating cytoskeletal function that contain multiple protein interaction domains (16). IQGAP1 is ubiquitously expressed and has been found to interact with APC (17) and the CLIP170 complex in response to small GTPases, promoting cell polarization and migration (18). IKKε is an IKK-related kinase that functions as part of the signal-stimulated noncanonical pathway of NF-kB activation (19). IKKε plays a role in the immune response and also impacts cell proliferation and transformation (20).
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