6-OHDA, apomorphine (APO), and ascorbic acid were purchased from Sigma Aldrich (St. Louis, MO, USA). INF-γ was bought from Sino Biological Inc. (Beijing, China). Fetal bovine serum (FBS) and Roswell Park Memorial Institute (RPMI) 1640 medium were obtained from HyClone (Logan, UT, USA) and Invitrogen (Carlsbad, CA, USA), respectively. Rabbit anti-TH antibody, rabbit anti-Iba1 antibody, and anti-GFAP antibody were obtained from Abcam (Cambridge, UK). Immunohistochemical kit (containing hydrogen peroxide, blocking solution, horseradish peroxidase- (HRP-) conjugated secondary antibody, and 3,3-diaminobenzidine (DAB)) was purchased from UNIV (Shanghai, China). SYBR Green Supermix was obtained from Bio-Rad (USA).
Rabbit anti iba1 antibody
Manufactured by Abcam
Sourced in United States, United Kingdom
The Rabbit anti-Iba1 antibody is a primary antibody that specifically recognizes the Iba1 (Ionized calcium-binding adapter molecule 1) protein. Iba1 is a protein involved in the activation and regulation of microglia, the immune cells of the central nervous system. This antibody can be used to detect and study the Iba1 protein in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti th antibody, by Abcam (2 mentions)
Mouse anti hsp70 antibody, by Abcam (2 mentions)
Sybr green supermix, by Bio-Rad (1 mentions)
Anti gfap antibody, by Abcam (1 mentions)
6 ohda, by Merck Group (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Apomorphine apo, by Merck Group (1 mentions)
Goat anti phosphorylated tau p tau, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti phf1 antibody, by Abcam (1 mentions)
Alexa fluor 488 or 594, by Jackson ImmunoResearch (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Goat anti gfap, by Abcam (1 mentions)
Mab1585, by Merck Group (1 mentions)
Rabbit anti iba1, by Abcam (1 mentions)
Goat anti vimentin, by Santa Cruz Biotechnology (1 mentions)
Goat anti gfap antibody, by Abcam (1 mentions)
Gapdh mouse monoclonal antibody, by Proteintech (1 mentions)
Rabbit anti nlrp3 antibody, by Abcam (1 mentions)
Hrp conjugated affinipure goat anti mouse rabbit igg h l, by Proteintech (1 mentions)
Rabbit anti myc tag antibody, by Abcam (1 mentions)
8 protocols using rabbit anti iba1 antibody
1
Neuroinflammation and Parkinson's Disease
2
Immunohistochemical Analysis of Alzheimer's Pathology
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Mouse hippocampi were formalin fixed and embedded in paraffin. Hematoxylin counterstaining was performed at the end. used to provide cytologic detail. Standard procedures of peroxidase-attached streptavidin and 3,3’-diaminobenzidine (DAB) as a substrate were sequentially used. Direct fluorescence was used to detect RFP. Immunostaining was done using rabbit anti-SPC25 antibody (Abcam, Dallas, TX, USA), rabbit anti-IBA-1 antibody (Abcam), rabbit anti-Aβ antibody (EMD Millipore, Billerica, MA, USA), or goat anti-phosphorylated Tau (p-Tau) (Santa Cruz Biotechnology, Dallas, TX, USA) or rabbit anti-PHF1 antibody (Abcam) antibodies. For quantitating neuronal cell loss, PHF-1+ neuronal positive cells were analyzed throughout the CA1 region of the hippocampus. Enzyme-linked immunosorbent assay (ELISA) for Aβ and pTau was performed using specific kits (R&D System, Los Angeles, CA, USA) as instructed.
3
Spinal Cord Injury Immunostaining
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Mice were perfused transcardially with ice-cold 4% paraformaldehyde in 0.1 M phosphate buffer (pH=7.2) at 0 and 14 days after injury. The spinal cords were removed and fixed in 4% paraformaldehyde. Fifteen-micrometer sections were prepared on a cryostat. Longitudinal section was made in order to show the structure of injured spinal cord well. The sections were blocked by using normal goat serum with 0.2% Triton-X 100 for 30 min at room temperature, and then incubated with Mouse Anti-HSP70 antibody (abcam) and Rabbit Anti-Iba1 antibody (abcam) overnight at 4oC and followed by staining with the corresponding secondary antibodies (Alexa Fluor 488 or 594, 1:800, Jackson Immuno Research) for 1 h at 37°C.
4
Quantification of M1 Microglia Activation
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After conventional dewaxing, coronal sections were incubated with rabbit anti-Iba-1 antibody (1:200, Abcam) or mouse anti-iNOS antibody (1:50, Santa Cruz Biotechnology, Dallas, TX, USA)
mixture overnight at 4°C. The sections were then treated with a Cy3-labeled goat anti-rabbit secondary antibody (1:200, Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA) and
FITC-labeled goat anti-mouse secondary antibody (1:200, Abcam) for 90 min at room temperature. Finally, sections were counterstained with DAPI (Aladdin Holdings Group Co., Ltd.) to stain
cellular nuclei. Fluorescent signals were detected under a fluorescence microscope (400× magnification, Olympus). Iba-1/iNOS-double-positive cells were considered M1 microglia, and the
percentages of M1 microglia in total cells (DAPI positive) and in microglia (Iba-1 positive) were calculated.
mixture overnight at 4°C. The sections were then treated with a Cy3-labeled goat anti-rabbit secondary antibody (1:200, Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA) and
FITC-labeled goat anti-mouse secondary antibody (1:200, Abcam) for 90 min at room temperature. Finally, sections were counterstained with DAPI (Aladdin Holdings Group Co., Ltd.) to stain
cellular nuclei. Fluorescent signals were detected under a fluorescence microscope (400× magnification, Olympus). Iba-1/iNOS-double-positive cells were considered M1 microglia, and the
percentages of M1 microglia in total cells (DAPI positive) and in microglia (Iba-1 positive) were calculated.
5
Midbrain Neuroinflammation Analysis
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After euthanizing the mice, their midbrains were removed and fixed using 4% paraformaldehyde. Then the midbrain was placed in 70%, 80%, 90%, 95%, and 100% alcohol for one hour each for dehydration. Then it was dipped in xylene for 30 min and immersed in wax for 60 min. The midbrain tissue was then sectioned (6 μm per slice) and dried. Dopaminergic neurons in SN were measured with rabbit anti-TH antibody (1:500, while microglia were detected with rabbit anti-IBA-1 antibody (1:500; Abcam, Cambridge, UK). The IHC staining was performed according to the instructions of the immunohistochemistry kit (Biological Technology, Wuhan, China). Finally, total TH and IBA-1-positive cells were counted by five researchers in a blinded fashion (who were unaware of the experimental design), and the average of these scores was reported. At least 3 slices per mouse were counted in the study.
6
Retinal Cell Labeling Protocol
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Two primary antibodies were used to label different populations of RGCs. The general population was labelled using a mouse anti-Brn3a antibody (1:500; mouse anti-Brn3a, MAB1585 Millipore, Madrid, Spain), and the ipRGCs were labelled with a rabbit anti-melanopsin antibody (1:500; rabbit anti-melanopsin; Invitrogen, Carlsbad, CA, USA). Microglial and macroglial (Müller cells and astrocytes) cells were labelled with rabbit anti-Iba 1 antibody (1:500; rabbit anti-Iba1; Abcam, USA, Waltham, MA, USA), goat anti-GFAP antibody (1:500; goat anti-GFAP; Abcam, USA) and goat anti-vimentin antibody (1:250, C-20, sc-7557; Santa Cruz Biotechnology, Dallas, TX, USA).
7
Comprehensive Antibody Validation Protocol
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Primary antibodies: rabbit anti-6×His tag antibody (Abcam), rabbit polyclonal anti-Tau antibody (Abcam), rabbit anti-Myc tag antibody (Abcam), mouse anti-Hsp70 antibody (Abcam), mouse anti-Hsp40 antibody (Abcam), mouse GAPDH monoclonal antibody (Proteintech), mouse anti-TNF-α antibody (ImmunoWay), rabbit anti-IL-18 antibody (ImmunoWay), rabbit anti-IL-1β antibody (ImmunoWay), rabbit anti-CAT antibody (Abcam), rabbit anti-GPX1 antibody (Abcam), rabbit anti-SOD1 antibody (Abcam), rabbit anti-ZsGreen antibody (Clontech), rabbit anti-ASC antibody (Cell Signaling Technology), rabbit anti-NLRP3 antibody (Abcam), rabbit anti-Iba1 antibody (Abcam), rabbit anti-GFAP antibody (Abcam). Secondary antibodies: HRP-conjugated Affinipure Goat Anti-Mouse/Rabbit IgG (H + L) (Proteintech), Alexa Fluor 488/594 (Invitrogen). All antibodies were diluted as the recommendation by the manufacturer.
8
Quantitative Analysis of Kidney Infiltrating Immune Cells
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Per our protocols, 25 (link) endogenous peroxidase was blocked with 2 % H 2 O 2 /methanol for 30 min and antigen retrieval in sodium citrate buffer (10 mM, pH 6.0) at 85 °C for 30 min. Sections were incubated with blocking buffer followed by primary antibodies at 4 °C overnight, including Rat anti-neutrophil monoclonal antibody (Thermo Scientific, Waltham, MA), rabbit anti-Iba1 antibody (Abcam, Waltham, MA), or rabbit anti-B220 antibody (Abcam), then incubated with biotinylated secondary antibodies for 2 hrs. The signal was visualized with 3,3′-diaminobenzidine (DAB). IgG control staining was performed following identical procedures excluding primary antibodies, which showed no positive signal (Fig. S5). Images were taken by a Nikon ECLIPSE E600 microscope with Nikon Elements software. Numbers of neutrophils, macrophages, and B cells (brown signal) and total cells (blue-purple hematoxylin stained nuclei) per unit area were counted (Supplementary Materials).
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