AIF-1/Iba1 Antibody

Immunocytochemistry/Immunofluorescence: AIF-1/Iba1 Antibody [NBP1-06014] - Immunofluorescence analysis of paraformaldehyde fixed Caco-2 cells, permeabilized with 0.15% Triton. Primary incubation 1hr using AIF-1/Iba1 Antibody (10ug/ml) followed by Alexa Fluor 488 secondary antibody (2 ug/mL), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 ug/mL) followed by Alexa Fluor 488 secondary antibody (2 ug/mL).

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody [NBP1-06014] - Staining of paraffin embedded Human Cerebellum AIF-1/Iba1 Antibody at 4 ug/mL. Microwaved antigen retrieval with Tris/EDTA buffer pH9, HRP-staining.

Flow Cytometry: AIF-1/Iba1 Antibody [NBP1-06014] - Flow cytometric analysis of paraformaldehyde fixed K562 cells (blue line), permeabilized with 0.5% Triton. Primary incubation 1hr (10 ug/mL) with AIF-1:Iba1 Antibody, followed by Alexa Fluor 488 secondary antibody (1 ug/mL). IgG control: Unimmunized goat IgG (black line) followed by Alexa Fluor 488 secondary antibody.

• Reactivity: Hu, Po, Bv, Ca, RM
• Applications: ELISA, Flow, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: 0.5 mg/ml

AIF-1/Iba1 Antibody Summary

• Immunogen: This AIF-1/Iba1 Antibody was developed against a peptide with sequence C-YEEKAREKEKP corresponding to C-Terminus according to NP_116573.1, NP_001614.3.
• Epitope: C-YEEKAREKEKP
• Marker: pan-Microglia Marker
• Specificity: This AIF-1/Iba1 Antibody is expected to recognise isoform 1 (NP_116573.1) and isoform 3 (NP_001614.3).
• Predicted Species: Porcine (91%), Bovine (91%), Rhesus Macaque (100%), Canine (100%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: AIF1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 10 ug/mL
  • Immunocytochemistry/ Immunofluorescence 10 ug/mL
  • Immunohistochemistry 2 - 4 ug/mL
  • Immunohistochemistry-Paraffin 2 - 4 ug/mL
  • Peptide ELISA Detection limit 1:16000
• Theoretical MW: 16.7 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.
• Control:

  AIF-1/Iba1 Overexpression Lysate

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: Tris saline (20 mM Tris pH 7.3, 150 mM NaCl), 0.5% BSA
• Preservative: 0.02% Sodium Azide
• Concentration: 0.5 mg/ml
• Purity: Immunogen 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 (NBP1-06014)(2)

Publications using NBP1-06014

• Zhu X, Schrader JM, Irizarry BA et al. Impact of A beta 40 and A beta 42 Fibrils on the Transcriptome of Primary Astrocytes and Microglia Biomedicines 2022-11-19 [PMID: 36428550] (IHC-Fr, Rat)
  
  Details:
  Fifteen-month-old rTgF344-AD rat brain sections, dilution used 1:200
• Kelemen SE, Autieri MV. Expression of allograft inflammatory factor-1 in T lymphocytes: a role in T-lymphocyte activation and proliferative arteriopathies. Am J Pathol 2005-08-01 [PMID: 16049345]

Bioinformatics

• Gene Symbol: AIF1
• Entrez:
  • 199(Human)
• Uniprot:
  • NP_116573.1(Human)