Mouse anti iba 1

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The mouse anti-Iba-1 antibody is a primary antibody used for the detection and localization of Iba-1 (Ionized calcium-binding adapter molecule 1), a protein that is specifically expressed in macrophages and microglia in the central nervous system. The antibody can be used in various applications, such as immunohistochemistry, immunocytochemistry, and western blotting, to study the distribution and function of Iba-1 in biological samples.

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Lab products found in correlation

Rabbit anti iba1, by Abcam (2 mentions) Alexa 594 conjugated goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions) Goat anti cd206, by R&D Systems (1 mentions) Quickblock, by Beyotime (1 mentions) Mouse anti cd86, by Santa Cruz Biotechnology (1 mentions) Rabbit anti trpv4, by Abcam (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Mouse anti neun antibody, by Merck Group (1 mentions) Alexa 594, by Thermo Fisher Scientific (1 mentions) Rabbit anti arginase 1, by Santa Cruz Biotechnology (1 mentions) Fv1000 confocal microscope, by Olympus (1 mentions) Mouse anti nos2, by Santa Cruz Biotechnology (1 mentions) N 1 naphthyl ethylenediamine dihydrochloride, by Merck Group (1 mentions) Goat anti mouse secondary antibody, by Abcam (1 mentions) Glutathione (gsh), by Merck Group (1 mentions) Phosphate buffered saline (pbs), by BD (1 mentions) 5 5 dithiobis 2 nitrobenzoic acid, by Merck Group (1 mentions) Formaldehyde, by BD (1 mentions) 1 chloro 2 4 dinitrobenzene, by Merck Group (1 mentions) Mounting media, by BD (1 mentions)

Close spelling variants of this product

Iba-1 (44 citations) Anti-Iba-1 (17 citations)

6 protocols using mouse anti iba 1

Immunofluorescence Staining of Brain Slices

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Brain slices were prepared as previously described,
²¹ (link) and 0.3% Triton ×100 was used to penetrate 15 min. Brain slices (30 μm) were blocked with QuickBlock™ (Beyotime, China) for 1 h, and then incubated in a refrigerator with primary antibodies at 4°C overnight. We performed single immunofluorescence staining using rabbit anti‐Iba1 (1:1000, Abcam, USA). Performed double immunofluorescence staining using mouse anti‐Iba1 (1:100, Santa Cruz Biotechnology, USA) with rabbit anti‐TRPV4 (1:200, Abcam, USA), rabbit anti‐Iba1 (1:1000, Abcam, USA) with mouse anti‐CD86 (1:50, Santa Cruz Biotechnology, USA), and rabbit anti‐Iba1 (1:1000, Abcam, USA) with goat anti‐CD206 (1:250, R&D Systems, USA). After washing the brain slices with PBS containing 0.3% Triton ×100, Alexa 594‐conjugated goat anti‐rabbit antibodies (1:1000, Life Technologies Corporation, USA), Alexa 555‐conjugated donkey anti‐goat antibodies (1:1000, Life Technologies Corporation, USA), and Alexa 488‐conjugated goat anti‐rabbit antibodies (1:1000, Life Technologies Corporation, USA) were added and incubated at room temperature for 2 h. The nucleus was stained with DAPI. Using laser scanning confocal microscopy (Leica SP‐8) to take pictures. ImageJ was used for analysis.

Ren X., Gao X., Li Z., Ding Y., Xu A., Du L., Yang Y., Wang D., Wang Z, & Shu S. (2024). Electroacupuncture ameliorates neuroinflammation by inhibiting TRPV4 channel in ischemic stroke. CNS Neuroscience & Therapeutics, 30(2), e14618.

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Assessing Paclitaxel-Induced Neuropathy Mitigation

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DRG tissues for assessing neuroprotective effect of minoxidil on paclitaxel-induced neuropathy were isolated at Day1, 4, or 7 after first paclitaxel injections. Sequentially, DRG tissues were fixed with 4% PFA for 4 hours and dehydrated with 30% sucrose in PBS overnight at 4 °C. For frozen sections, the samples were embedded in OCT (Leica, IL, USA) and sliced into 20 μm thick sections. These DRG tissue sections were washed with 0.05% Triton X-100 in PBS for 30 minutes and blocked with 3% BSA at room temperature for 1 hour. DRG tissue samples were co-stained with mouse anti-NeuN antibody (1:100; Millipore), mouse anti-NOS2 (1:100; Santa Cruz), mouse anti-Iba-1 (1:100; Santa Cruz), rabbit anti-Arginase-1 (1:100; Santa Cruz), rabbit anti-Iba-1 (1:100; Abcam) and rat anti-CD68 antibody (1:100; Novus, USA) at 4 °C overnight. Then, these samples were washed by PBS and incubated in secondary antibodies (Alexa-488 and Alexa-594 1:200; Invitrogen, CA) at room temperature for 1 hour. Immunofluorescence images of DRG tissues were acquired using a 40x objective by Olympus FV1000 confocal microscope (Olympus, Japan).

Chen Y.F., Chen L.H., Yeh Y.M., Wu P.Y., Chen Y.F., Chang L.Y., Chang J.Y, & Shen M.R. (2017). Minoxidil is a potential neuroprotective drug for paclitaxel-induced peripheral neuropathy. Scientific Reports, 7, 45366.

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Anti-inflammatory Compound Characterization

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All the starting materials were purchased from Daejung (South Korea) and Alfa-Aesar (Germany). Digital Gallenkamp (Sanyo) apparatus was used to record the melting points of final compounds and was uncorrected. Proton NMR (¹H-NMR) and carbon-13 (¹³C-NMR) spectra were checked using Bruker AM-300 in DMSO-d6 at 300 MHz and 75 MHz, respectively, while using TMS as an internal standard. Alpha Bruker FTIR spectrophotometer (ATR eco ZnSe, νmax in cm⁻¹) was used to record FTIR spectra. All reactions were monitored by thin-layer chromatography (TLC). 3,3-diaminobenzidine peroxidase, an ABC Elite kit, mouse anti-COX-2, mouse anti-p-NF-κB, and mouse anti-Iba-1 were procured from (Santa Cruz Biotechnology, Dallas, TX, USA). H₂O₂, Proteinase K, formaldehyde, mounting media, and PBS tablets were acquired from BDH while secondary antibody goat anti-mouse was purchased from Abcam (Cambridge, UK). Other chemicals were procured from Sigma-Aldrich (St. Louis, MO, USA) like solvents and reagents such as 5,5′-dithiobis (2-nitrobenzoic acid), 1-chloro-2,4-dinitrobenzene, glutathione (GSH), N-(1-naphthyl) ethylenediamine dihydrochloride, and trichloroacetic acid. All chemicals used were of high analytical grade (99% HPLC).

Imran M., Al Kury L.T., Nadeem H., Shah F.A., Abbas M., Naz S., Khan A.U, & Li S. (2020). Benzimidazole Containing Acetamide Derivatives Attenuate Neuroinflammation and Oxidative Stress in Ethanol-Induced Neurodegeneration. Biomolecules, 10(1), 108.

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Immunocytochemistry for Microglial Morphology

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Morphological changes in microglia were determined via immunocytochemistry using commercially available antibodies against IBA-1. Unstimulated and LPS-stimulated (for 24 h) microglia were cultured on sterile cover slips in 6-well plates (1 × 10⁶ cells/well). The cells were fixed for 20 min in 4% paraformaldehyde (Sigma-Aldrich, St. Louis, MO, United States). Then, the cells were permeabilized with 0.1% Triton^TM X-100 (Sigma-Aldrich, St. Louis, MO, United States) in PBS for 30 min at room temperature, washed with PBS, and blocked with 5% donkey serum in PBS. The microglial cells were incubated in primary antibodies [mouse anti-IBA-1 (Santa Cruz Biotechnology Inc., United States)], overnight at 4°C. After washing with PBS, the microglia were incubated for 2–3 h in a fluorophore-conjugated secondary antibody: Alexa Fluor 546 donkey anti-mouse (Molecular Probes) diluted 1:500 in 5% normal donkey serum (NDS). Then, the cells were washed with PBS and coverslipped using Aquatex mounting medium (Merck, Darmstadt, Germany). Morphological changes after LPS administration were evaluated by visualizing microglia under the 40× objective of a Zeiss microscope (Zeiss, Germany). Sections without primary antibodies were used as negative controls.

Rojewska E., Ciapała K., Piotrowska A., Makuch W, & Mika J. (2018). Pharmacological Inhibition of Indoleamine 2,3-Dioxygenase-2 and Kynurenine 3-Monooxygenase, Enzymes of the Kynurenine Pathway, Significantly Diminishes Neuropathic Pain in a Rat Model. Frontiers in Pharmacology, 9, 724.

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Immunohistochemical Analysis of Microglial Activation

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At 24 h or 72 h after surgery, animals were deeply anaesthetized and perfused with saline, followed by 4% paraformaldehyde. The brains were gathered and dehydrated in 30% sucrose solution overnight. After that, the brains were placed in OCT and cut into 20 μm frozen segments. The slices were placed in blocking buffer in 8% goat serum for 2 h at room temperature and then treated with a primary antibody: mouse anti-Iba1 (1 : 100, Santa Cruz Biotechnology, USA), rabbit anti-CD16 (1 : 100, Abcam, USA), and rabbit anti-CD206 (1 : 100, Abcam, USA) at 4°C overnight. Membranes were washed with PBST three times (0.1% tween 20 in PBS); the tissue sections were treated with FITC- or Cy3-conjugated secondary antibody (1 : 100, BIOSS, China) and at room temperature for 2 h. Finally, nuclei were costained with DAPI (Beyotime Institute of Biotechnology, China) for 30 min. Images were obtained through a laser scanning confocal microscope (LSM700, Carl Zeiss, Germany) and assessed using ImageJ software. The stained cells were counted in the ipsilateral cortex penumbra under 400x magnification.

Han D., Wang J., Wen L., Sun M., Liu H, & Gao Y. (2021). Remote Limb Ischemic Postconditioning Protects against Ischemic Stroke via Modulating Microglia/Macrophage Polarization in Mice. Journal of Immunology Research, 2021, 6688053.

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Microglial Activation Assessed by IBA1 and CD68

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To confirm microglial lineage and the effect of the different treatments on its activation, we assessed the expression of ionized calcium-binding adapter molecule 1 (IBA1) and CD68 by immunofluorescence analyses.
Microglial cells cultured for 1 day on permanox chamber slide at the concentration of 2.5 • 10 4 cell/cm 2 were treated with LPS alone or in combination with ADSC-CM. After 48 h at 37°C, the cells were fixed with 4% PFA (15 min, RT). To avoid quenching, microglia were treated with 0.1 M glycine (10 min, RT). Subsequently, the samples were permeabilized with 0.1% Triton X-100 for 15 min. The following primary antibodies were applied overnight at 4°C: mouse anti-IBA1 (1:500) and mouse anti-CD68 (1:400), (both purchased from Santa Cruz Biotechnology, Inc., Heidelberg, Germany). Cells were then incubated with Alexa488 conjugated secondary antibody (1:1,000; Life Technologies) at RT for 45 min, washed, and the nuclei were counterstained with DAPI (Life Technologies). The sample was mounted with ProLong Ò Gold Antifade Mountant (Life Technologies) and images were captured using a Leica TCS SP5 confocal microscope (Leica Microsystems, Wetzlar, Germany). Cells were counted in three microscopic fields in each well (three wells/treatment) and expressed as a percentage of the total number of cells. Each treatment was performed in triplicate.

Marfia G., Navone S.E., Hadi L.A., Paroni M., Berno V., Beretta M., Gualtierotti R., Ingegnoli F., Levi V., Miozzo M., Geginat J., Fassina L., Rampini P., Tremolada C., Riboni L, & Campanella R. (2016). The Adipose Mesenchymal Stem Cell Secretome Inhibits Inflammatory Responses of Microglia: Evidence for an Involvement of Sphingosine-1-Phosphate Signalling. Stem cells and development, 25(14).

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