Cells were gently collected and fixed for 30 min with ice-cold 4% paraformaldehyde. When necessary, cells were permeabilized with 0.1% Triton X-100 for 10 min on ice. After blocking with 3% BSA for 30 min, cells were incubated with primary antibodies overnight at 4 °C, then washed prior to incubation with secondary antibodies for 45 min at RT. Primary antibodies used were: chicken anti-integrin α-M (1:500, Aves), mouse anti-GFAP (1:500, Cell Signaling), rabbit anti-BDNF (1:500; S. Cruz), goat anti-TNFα (1:250; S. Cruz), rabbit anti-Iba1 (1:300; Novus Biologicals, NBP2-19019). Antibodies were appropriately combined in multiple labeling experiments. Secondary antibodies used were: PE-anti-chicken (1:200; S. Cruz), FITC-anti-goat (1:200; S. Cruz sc-2024), Alexa-Fluor 647-anti-rabbit (1:500; Invitrogen A31573), PE-anti-mouse (1:300; S. Cruz sc-3738). Data was acquired on the Amnis FlowSight® Imaging Flow Cytometer and analyzed with IDEAS® Software (Millipore).
Rabbit anti iba1
Manufactured by Novus Biologicals
Rabbit anti-Iba1 is a primary antibody that specifically binds to the Iba1 (Ionized calcium-binding adapter molecule 1) protein. Iba1 is a calcium-binding protein that is expressed in macrophages and microglia, and it is commonly used as a marker for these cell types. The Rabbit anti-Iba1 antibody can be used in various immunodetection techniques, such as western blotting, immunohistochemistry, and immunocytochemistry, to identify and visualize Iba1-positive cells.
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Lab products found in correlation
2 protocols using rabbit anti iba1
1
Multimarker Immunophenotyping of Fixed Cells
2
Immunoblot Analysis of Spinal Cord Lesions
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NR-labeled lesion and unlabeled nonlesion sites were dissected from spinal cords at 5, 10, and 20 dpl as described above. The tissue was lysed in RIPA buffer (Millipore), separated by SDS/PAGE, and immunoblotted using the following antibodies: rabbit anti-Iba1 (1:500; Novus), rabbit anti-MOG (1:200; Santa Cruz Biotechnology), and rabbit β-actin (1:5,000 Abcam) for loading control. Proteins were detected using horseradish peroxidase-conjugated secondary antibodies and Pierce ECL Western blotting substrate.
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