AIF-1/Iba1 Antibody (JM36-62)

Western Blot: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Analysis of Iba1 on THP-1 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (1:500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:5000 dilution was used for 1 hour at room temperature.

Immunocytochemistry/Immunofluorescence: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Staining AIF-1/Iba1 in SH-SY5Y cells using (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X-100, PBS.

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Analysis of paraffin-embedded rat lung tissue using AIF-1/Iba1 Antibody (JM36-62). Counterstained with hematoxylin.

Flow Cytometry: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Analysis of THP-1 cells withAIF-1:Iba1 Antibody (JM36-62) at 1:100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black).

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Analysis of paraffin-embedded human lung cancer tissue using AIF-1/Iba1 Antibody (JM36-62). Counterstained with hematoxylin.

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Analysis of paraffin-embedded human spleen tissue using AIF-1/Iba1 Antibody (JM36-62). Counterstained with hematoxylin.

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Analysis of paraffin-embedded mouse brain tissue using AIF-1/Iba1 Antibody (JM36-62). Counterstained with hematoxylin.

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Analysis of paraffin-embedded mouse spleen tissue using AIF-1/Iba1 Antibody (JM36-62). Counterstained with hematoxylin.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Flow, ICC/IF, IHC, IHC-P, IP
• Clone: JM36-62
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

AIF-1/Iba1 Antibody (JM36-62) Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Recombinant AIF-1/Iba1 protein
• Localization: Cytoplasm, cytoskeleton.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: AIF1
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:500-1:1000
  • Flow Cytometry 1:50-1:200
  • Immunocytochemistry/Immunofluorescence 1:50-1:100
  • Immunohistochemistry 1:100-1:500
  • Immunohistochemistry-Paraffin 1:100-1:500
  • Immunoprecipitation
• Application Notes: IP: Use at an assay dependent concentration.
• 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.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: TBS (pH7.4), 1% BSA, 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for AIF-1/Iba1 Antibody (JM36-62)

• 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 INF-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.

Bioinformatics

• Gene Symbol: AIF1