For the assessment of number of activated microglia, brain sections were immunostained with anti-ionized calcium binding adaptor molecule-1. (Iba-1(ab5076); 1:500; Abcam, Tokyo, Japan) as previously described [26 (link)]. The number of positive cells was counted in both the CA1 region of hippocampus (1 field was counted in each side of CA1 region at × 200 magnification) and the cortex (2 fields were counted in each side at × 200 magnification). The number of cells was expressed as cells/mm2.
Iba 1 ab5076
Manufactured by Abcam
Sourced in Japan, United Kingdom
Iba-1(ab5076) is a polyclonal antibody that recognizes the Iba-1 protein. Iba-1 is a calcium-binding protein that is specifically expressed in macrophages and microglia. The antibody can be used for the detection of Iba-1 in various applications, such as Western blotting and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Emd638683, by MedChemExpress (1 mentions)
Gfap g3893, by Merck Group (1 mentions)
Bim sc 374358, by Santa Cruz Biotechnology (1 mentions)
Artemisinin, by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Tnf α, by Dakewe (1 mentions)
Bv2 microglial cells, by Jennio Biotech (1 mentions)
Il 1β, by Dakewe (1 mentions)
Mcp 1, by Dakewe (1 mentions)
4 protocols using iba 1 ab5076
1
Quantifying Activated Microglia in Brain Sections
2
Immunohistochemical Staining of Iba-1 in Tissue Sections
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The tissues were dehydrated in a graded series of ethanol, cleared in xylene and embedded in paraffin blocks. The blocks of tissue were sectioned serially at 5 μm using a microtome. After the tissues were mounted on the slide, deparaffinated and rehydrated, they were immersed in sodium citrate for antigen retrieval using a microwave oven (medium power for 6 min, 4 times). The sections were treated with 3% H2O2 for 10 min and washed three times with PBS. These sections were blocked with 5% bovine serum albumin (BSA) in PBS for 30 min and incubated overnight with the primary antibody Iba-1 (ab5076, 1:400, Abcam) at 4°C. After three PBS washes, these sections were incubated with biotinylated-conjugated rabbit anti-goat IgG secondary antibody for 30 min at 37°C. After three PBS washes, sections were treated with SABC for 30 min at 37°C and then washed three times with PBS. A DAB kit was used for chromogenic detection, and the sections were then stained with haematoxylin and mounted. The images were captured by an OLYMPUS microscope.
3
Neuroinflammation and Neurogenesis Assays
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Antibodies against Bcl-2 interacting mediator of cell death (Bim;sc-374358) and B-cell lymphoma-extra large were obtained from Santa Cruz Biotechnology (Santa Cruz, CA); anti-parvalbumin (PV;P3088) and anti-glial fibrillary acidic protein (GFAP;G3893) antibodies were obtained from Sigma-Aldrich; bromodeoxyuridine (BrdU, NBP2-14890) was obtained from Novus; rabbit Anti-Goat IgG conjugated to Alexa Fluor® 488 (ab150141), anti-Doublecortin (DCX;ab153668) and anti-ionized calcium-binding adapter molecule 1 (Iba-1;ab5076) were obtained from Abcam; goat anti-mouse IgG conjugated to Alexa Fluor® 594 (A-11005), goat anti-chicken IgY conjutated to Alexa Fluor® 594 (A-11042), goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (A-11008) and anti-calcium/calmodulin-dependent protein kinase II (CaMKII;MA1-048) were obtained from Thermo Fisher Scientific. A rabbit polyclonal antibody against SGK1.1 was generated against a glutathione Stransferase (GST) fusion protein containing amino acids 41-80 from the SGK1.1-specific exon 2, expressed in E. coli and purified by affinity chromatography. KA (K0250) and M-current blocker XE991 (X2254) were obtained from Sigma Aldrich. SGK1 inhibitor EMD638683 was from MedChem Express (HY-15193A).
4
Artemisinin Modulates Microglial Activation
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BV2 microglial cells were purchased from Guangzhou Jennio Biotech Co., Ltd (Guangzhou, China). Artemisinin (purity > 98%), LPS and DMSO were from Sigma Aldrich (MO, USA). Iba-1 (ab5076) and TNF-α (ab1793) antibodies were purchased from Abcam (Cambridge, UK). AMPKα1 (cst 2795s), p-AMPKα1 (cst 4185s), iNOS (cst 13120s), IL-6 (cst 12912), NF-κB (cst 8242s), PCNA (cst 13100s), α-Tubulin (cst 2125s) and β-actin (cst 4970s) antibodies were purchased from Cell Signaling Technology Inc (Danvers, USA).
Cleaved caspase 3 (GB11532) and CD68 (GB11067) antibodies was purchased from Wuhan Servicebio Biotechnology, Co., Ltd. (Wuhan, China). P2Y12 antibody (PAB32958) was purchased from Biowamp Life Science Lab, Co., Ltd. (Wuhan, China). TNF-α, IL-6, IL-1α, IL-1β, MCP-1 and MIP-2 ELISA kits were purchased from Dakewe Bio-engineering Co., Ltd (Shenzhen, China).
Cleaved caspase 3 (GB11532) and CD68 (GB11067) antibodies was purchased from Wuhan Servicebio Biotechnology, Co., Ltd. (Wuhan, China). P2Y12 antibody (PAB32958) was purchased from Biowamp Life Science Lab, Co., Ltd. (Wuhan, China). TNF-α, IL-6, IL-1α, IL-1β, MCP-1 and MIP-2 ELISA kits were purchased from Dakewe Bio-engineering Co., Ltd (Shenzhen, China).
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