AIF-1/Iba1 Antibody (JM36-62)

Novus Biologicals | Catalog # NBP2-75397

Recombinant Monoclonal Antibody
Novus Biologicals
100% Guarantee

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse, Rat

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # JM36-62 expressed in HEK293
View all AIF-1/Iba1 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide within N-terminal human AIF-1/Iba1. (SwissProt: P55008 Human; SwissProt: O70200 Mouse; SwissProt: P55009 Rat)

Localization

Cell membrane, Cell projection, Cytoplasm, Cytoskeleton, Membrane.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

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.

Scientific Data Images for AIF-1/Iba1 Antibody (JM36-62)

Western Blot: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Western Blot: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

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]

Immunocytochemistry/ Immunofluorescence: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

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]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Immunofluorescence analysis of paraffin-embedded rat brain tissue labeling Iba1 with Rabbit anti-AIF-1/Iba1 antibody washed with PBS, and then probed with the primary antibody (green) at 1/100 dilution overnight at 4, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor(TM) 488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
Flow Cytometry: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Flow Cytometry: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

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]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

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]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

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]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

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]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

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.
Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

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.
Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Immunofluorescence staining of paraffin- embedded human spleen tissue using anti-AIF-1/Iba1rabbit polyclonal antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.(sodium citrate buffer (pH6) for 20 mins.) The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with at 1/50 dilution for 10 hours at 4C and detected using Alexa Fluor(R) 488 conjugate-Goat anti-Rabbit IgG (H+L) Secondary Antibody at a dilution of 1:500 for 1 hour at room temperature.
Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Immunofluorescence staining of paraffin- embedded mouse brain tissue using anti-AIF-1/Iba1 antibody.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with at 1/50 dilution for 10 hours at 4 and detected using Alexa Fluor(R) 488 conjugate-Goat anti-Rabbit IgG (H+L) Secondary Antibody at a dilution of 1:500 for 1 hour at room temperature.
Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-AIF-1/Iba1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Analysis of paraffin-embedded human kidney tissue with Rabbit anti-AIF-1/Iba1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Analysis of paraffin-embedded human spleen tissue with Rabbit anti-AIF-1/Iba1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397]

Immunohistochemistry-Paraffin: AIF-1/Iba1 Antibody (JM36-62) [NBP2-75397] - Analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Iba1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Applications for AIF-1/Iba1 Antibody (JM36-62)

Application
Recommended Usage

Flow Cytometry

1:50-1:200

Immunocytochemistry/ Immunofluorescence

1:50-1:100

Immunohistochemistry

1:10 - 1:500

Immunohistochemistry-Frozen

1:50

Immunohistochemistry-Paraffin

1:100-1:500

Immunoprecipitation

1:10 - 1:500

Western Blot

1:500-1:1000
Application Notes
IP: Use at an assay dependent concentration.

Reviewed Applications

Read 1 review rated 1 using NBP2-75397 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

TBS (pH7.4), 0.05% BSA, 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: AIF-1/Iba1

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

Long Name

Allograft Inflammatory Factor 1

Alternate Names

AIF1, IBA1, IRT1

Gene Symbol

AIF1

Additional AIF-1/Iba1 Products

Product Documents for AIF-1/Iba1 Antibody (JM36-62)

Certificate of Analysis

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Product Specific Notices for AIF-1/Iba1 Antibody (JM36-62)

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.

Citations for AIF-1/Iba1 Antibody (JM36-62)

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  • AIF-1/Iba1 Antibody (JM36-62)
    Name: Yuanxia Wu
    Application: Immunohistochemistry-Frozen
    Sample Tested: Rat hippocampus
    Species: Rat
    Verified Customer | Posted 01/06/2021
    No microglial cell was observed.
    AIF-1/Iba1 Antibody (JM36-62) NBP2-75397
    Bio-Techne Response
    This review was submitted through the legacy Novus Innovators Program, reflecting a new species or application tested on a primary antibody.

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