AIF-1/Iba1 Antibody

Immunoprecipitation: AIF-1/Iba1 Antibody [NBP2-16908] - Immunoprecipitation of AIF-1/Iba1 protein from K562 whole cell extracts using 5ug of AIF-1/Iba1 Antibody Western blot analysis was performed using Iba1 antibody EasyBlot anti-Rabbit IgG was used as a secondary reagent.

Immunocytochemistry/Immunofluorescence: AIF-1/Iba1 Antibody [NBP2-16908] - THP-1 cells were fixed in 4% paraformaldehyde at RT for 15 min.Green: Iba1 stained by Iba1 antibody diluted at 1:500.Blue: Fluoroshield with DAPI .

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NBP2-16908] - Rat brain. Iba1 stained by Iba1 antibody diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Flow Cytometry: AIF-1/Iba1 Antibody [NBP2-16908] - THP-1 cell. Black: Unlabelled sample was used as a control. Red: AIF-1:Iba1 Antibody dilution: 1:50. Acquisition of 20,000 events were collected using a Dylight 488-conjugated secondary antibody for FACS analysis.

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NBP2-16908] - Mouse cerebellum. Iba1 stained by Iba1 antibody diluted at 1:1000. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NBP2-16908] - Rat cerebellum. Iba1 stained by Iba1 antibody diluted at 1:1000.Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NBP2-16908] - Mouse brain. Iba1 stained by Iba1 antibody diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NBP2-16908] - Mouse brain. Iba1 stained by Iba1 antibody diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NBP2-16908] - Rat brain. Iba1 stained by Iba1 antibody diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NBP2-16908] - Mouse brain. Iba1 stained by Iba1 antibody diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Flow Cytometry: AIF-1/Iba1 Antibody [NBP2-16908] - Primary murine microglia cells, staining with AIF-1:Iba1 Antibody using 1. 0 ug per 4 x 10^5 cells. NBP2-16908 (blue), Rabbit IgG (green) Unstained (red).

Western Blot: AIF-1/Iba1 Antibody [NBP2-16908] - Various tissue extracts (50 ug) were separated by 15% SDS-PAGE, and the membrane was blotted with AIF-1/Iba1 antibody (NBP2-16908) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Western Blot: AIF-1/Iba1 Antibody [NBP2-16908] - Various whole cell extracts (30 ug) were separated by 15% SDS-PAGE, and the membrane was blotted with AIF-1/Iba1 antibody (NBP2-16908) diluted at 1:5000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

• Reactivity: Hu, Mu, Rt, Po, RM
• Applications: WB, Flow, ICC/IF, IHC, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

AIF-1/Iba1 Antibody Summary

• Immunogen: Recombinant protein encompassing a sequence within the center region of human AIF-1/Iba1. The exact sequence is proprietary.
• Marker: pan-Microglia Marker
• Predicted Species: Porcine (93%), Rhesus Macaque (99%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: AIF1
• Purity: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:50-1:200
  • Immunocytochemistry/ Immunofluorescence 1:100-1:1000
  • Immunohistochemistry 1:100-1:1000
  • Immunohistochemistry-Frozen 1:100-1:1000
  • Immunohistochemistry-Paraffin 1:100-1:1000
  • Immunoprecipitation 1:100-1:500
  • Western Blot 1:500-1:10000
• Theoretical MW: 17 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Bovine (88%).

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS, 1% BSA, 20% Glycerol
• Preservative: 0.025% Proclin 300
• Purity: Antigen Affinity-purified

Alternate Names for AIF-1/Iba1 Antibody

• AIF1
• AIF-1
• allograft inflammatory factor 1
• G1
• IBA1
• IbaI
• interferon gamma responsive transcript
• Ionized calcium-binding adapter molecule 1
• IRT1
• IRT-1
• Protein G1

Background

Allograft inflammatory factor (AIF-1) or ionized calcium-binding adapter molecule 1 (Iba1) is a cytosolic actin binding protein containing a calcium binding domain (EF-hand) and inducible by cytokines and IFN-gamma (1). AIF-1 (theoretical molecular weight 17kDa) was first cloned from rat and human cardiac allografts and macrophage cell lines. In cardiac allografts, expression of AIF-1 was specifically associated with infiltrated mononuclear cells (2, 3). AIF-1 is constitutively expressed and inducible in macrophages and microglia. Iba1, microglia response factor (MRF-1) and daintain were independently cloned and identified in rat and human tissues and share complete sequence identity with AIF-1. Additionally, several AIF-1 splice variants have been identified including IRT-1, BART-1, G1 and Hara-1 (2).

Several cellular functions have been associated with AIF-1/Iba1 expression including cell growth, cell migration, actin bundling, membrane ruffling, and phagocytic activity (2, 4). Iba1 induces Rac signaling through a PLC-gamma dependent pathway (1). Rac, a member of the Rho family of small GTPases, localizes with Iba1 and F-actin in membrane ruffles and phagocytic cups and plays a role in microglia activation. AIF-1/Iba1 induction in macrophages and microglia occur in association with immunological inflammatory processes in various disease states including endometriosis, cerebral infarction and rheumatoid arthritis (5). Immunodetection of Iba1 through flow cytometry, immunohistochemical or immunocytochemical applications is commonly used for identification and analysis of microglia.

References

1. Imai, Y., & Kohsaka, S. (2002). Intracellular signaling in M-CSF-induced microglia activation: Role of Iba1. GLIA. https://doi.org/10.1002/glia.10149

2. Deininger, M. H., Meyermann, R., & Schluesener, H. J. (2002). The allograft inflammatory factor-1 family of proteins. FEBS Letters. https://doi.org/10.1016/S0014-5793(02)02430-4

3. Utans, U., Quist, W. C., Mcmanus, B. M., Wilson, J. E., Arceci, R. J., Wallace, A. F., & Russell, M. E. (1996). Allograft inflammatory factor-1: A cytokine-responsive macrophage molecule expressed in transplanted human hearts. Transplantation. https://doi.org/10.1097/00007890-199605150-00018

4. Franco-Bocanegra, McAuley, Nicoll, & Boche. (2019). Molecular Mechanisms of Microglial Motility: Changes in Ageing and Alzheimer's Disease. Cells. https://doi.org/10.3390/cells8060639

5. Kimura, M., Kawahito, Y., Obayashi, H., Ohta, M., Hara, H., Adachi, T., ... Yoshikawa, T. (2007). A Critical Role for Allograft Inflammatory Factor-1 in the Pathogenesis of Rheumatoid Arthritis. The Journal of Immunology. https://doi.org/10.4049/jimmunol.178.5.3316

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for AIF-1/Iba1 Antibody (NBP2-16908)(7)

Publications using NBP2-16908

• Pasula MB, Sylvester PW, Briski KP., et Al. Sex-dimorphic glucose transporter-2 regulation of cAMP-protein kinase A (PKA) C-alpha pathway activity and phosphorylation in rat hypothalamic primary astrocyte cultures Eur J Neurosci 2024-11-26 [PMID: 39592424]
• An SSA, Shim KH, Kang S Et al. The potential anti-amyloidogenic candidate, SPA1413, for Alzheimer's disease British journal of pharmacology 2021-10-05 [PMID: 34610141]
• Yeo IJ, Yun J, Son DJ et al. Antifungal drug miconazole ameliorated memory deficits in a mouse model of LPS-induced memory loss through targeting iNOS Cell Death Dis 2020-08-14 [PMID: 32796824] (WB, Mouse)
• Gimenes AD, Andrade BFD, Pinotti JVP et al. Annexin A1-derived peptide Ac2-26 in a pilocarpine-induced status epilepticus model: anti-inflammatory and neuroprotective effects J Neuroinflammation. 2019-02-11 [PMID: 30755225] (WB, Rat)
• Sun X, Bai Y, Zhai H et al. Devising micro/nano-architectures in multi-channel nerve conduits towards a pro-regenerative matrix for the repair of spinal cord injury Acta Biomater 2018-12-23 [PMID: 30586646] (ICC/IF, Rat)
• Schultz Mk, Gentzel R, Usenovic M et al. Pharmacogenetic neuronal stimulation increases human tau pathology and trans-synaptic spread of tau to distal brain regions in mice. Neurobiol. Dis. 2018-07-07 [PMID: 30049665] (IF/IHC, Mouse)
• Chisari M, Scuderi A, Ciranna L et al. Purinergic P2Y1 Receptors Control Rapid Expression of Plasma Membrane Processes in Hippocampal Astrocytes. Mol. Neurobiol. 2016-06-18 [PMID: 27318677] (ICC/IF, Rat)

Bioinformatics

• Gene Symbol: AIF1