Anti-Iba1 antibody [EPR16589] is a rabbit recombinant monoclonal antibody that is used to detect Iba1 in ICC/IF, IHC (PFA fixed), IHC-Fr, IHC-P, IP, Western blot. Suitable for Human, Mouse, Rat samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Antibody clone EPR16589 is cited in over 370 publications
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IHC-Fr | IP | WB | ICC/IF | IHC (PFA fixed) | IHC-P | |
---|---|---|---|---|---|---|
Human | Expected | Expected | Tested | Tested | Expected | Tested |
Mouse | Tested | Tested | Tested | Expected | Tested | Tested |
Rat | Tested | Expected | Tested | Expected | Expected | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/200 | Notes - |
Species Rat | Dilution info 1/200 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/40 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/8000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/8000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/8000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Actin-binding protein that enhances membrane ruffling and RAC activation. Enhances the actin-bundling activity of LCP1. Binds calcium. Plays a role in RAC signaling and in phagocytosis. May play a role in macrophage activation and function. Promotes the proliferation of vascular smooth muscle cells and of T-lymphocytes. Enhances lymphocyte migration. Plays a role in vascular inflammation.
G1, IBA1, AIF1, Allograft inflammatory factor 1, AIF-1, Ionized calcium-binding adapter molecule 1, Protein G1
Anti-Iba1 antibody [EPR16589] is a rabbit recombinant monoclonal antibody that is used to detect Iba1 in ICC/IF, IHC (PFA fixed), IHC-Fr, IHC-P, IP, Western blot. Suitable for Human, Mouse, Rat samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Antibody clone EPR16589 is cited in over 370 publications
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Find all reagents to label astrocytes in our "Microglia markers guide".
Anti-Iba1 antibody [EPR16589] (ab178847) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in ICC/IF, IHC (PFA fixed), IHC-P, IP and WB.
Anti-Iba1 antibody [EPR16589] (ab178847) was first used in a scientific publication in 2016 and has been cited over 284 times in peer reviewed journals. It's performance in immunofluorescence and IHC in mouse and rat samples is trusted by the scientific community.
Abcam's high quality manufacturing and validation processes ensure Anti-Iba1 antibody [EPR16589] (ab178847) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
Anti-Iba1 antibody [EPR16589] (ab178847) has 18 independent reviews from customers.
Anti-Iba1 antibody [EPR16589] (ab178847) specifically detects Iba1 (UniProt ID: P55008; Molecular weight: 17kDa) and is sold in 100 µL and 1 mL selling sizes.
Conjugation-ready, carrier free format available for antibody clone EPR16589 - Anti-Iba1 antibody [EPR16589] - BSA and Azide free ab221790.
Antibody clone EPR16589 is also available pre-conjugated to a variety of labels for your convenience - HRP, Alexa Fluor® 488, Alexa Fluor® 647, Alexa Fluor® 594, Alexa Fluor® 488, Alexa Fluor® 568, Alexa Fluor® 555, Alexa Fluor® 750 (HRP Anti-Iba1 antibody [EPR16589] ab216544, Alexa Fluor® 488 Anti-Iba1 antibody [EPR16589] ab310987, Alexa Fluor® 647 Anti-Iba1 antibody [EPR16589] ab311112, Alexa Fluor® 594 Anti-Iba1 antibody [EPR16589] ab311734, Alexa Fluor® 488 Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (Chimeric) ab312913, Alexa Fluor® 568 Anti-Iba1 antibody [EPR16589] ab313011, Alexa Fluor® 555 Anti-Iba1 antibody [EPR16589] ab313215, Alexa Fluor® 750 Anti-Iba1 antibody [EPR16589] ab321219).
NEW: Explore our alternative host species for clone [EPR16589] : rat, mouse. Iba1 (AIF1) is a marker for microglia, the brain's primary immune cells and is frequently used in Alzheimer's disease (AD) research. In AD brains, altered Iba1 expression indicates microglial activation and neuroinflammation. Researchers use Iba1 to assess microglial response and motility in AD models, making it essential for understanding the role of inflammation and immune response in Alzheimer's pathology.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Iba1 also known as Allograft Inflammatory Factor 1 (AIF-1) is a 17 kDa protein notable for its involvement in immune responses. It is expressed mainly in microglia and macrophages key components of the central nervous system's immune system. Researchers often use Iba1 in Western blots and immunohistochemistry (IHC) to study microglial activation. The protein's expression is a strong marker of microglial activation making it a focal point in neuroinflammation studies. Anti-Iba1 antibodies have gained popularity for their effectiveness in identifying microglia in brain tissue sections through Iba1 staining techniques.
Iba1 modulates actin bundling facilitating changes in microglial morphology and motility. It does not form part of a multiprotein complex but is significantly involved in cytoskeletal dynamics. This protein's expression increases during brain injury indicating its role in response to neural trauma. Iba1 has calcium-binding properties which suggest that it might interact with proteins involved in calcium signaling pathways.
Calcium signaling and actin cytoskeletal rearrangement are significant biological processes that involve Iba1. Within these pathways Iba1 interacts closely with other actin-binding proteins that affect microglial movement and morphology. Such interactions illustrate how Iba1 helps microglia navigate and respond to changes within the brain environment.
Iba1 is often associated with neurodegenerative diseases like Alzheimer's disease and amyotrophic lateral sclerosis (ALS). In these conditions elevated Iba1 levels mark increased microglial activity which correlates with disease progression. Connections with proteins such as amyloid-beta in Alzheimer's disease suggest its role in mediating inflammatory responses and linking inflammation to disease pathology.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
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Terms & Conditions.
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Iba1 with ab178847 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Cytoplasm staining on microglia of the rat cerebrum is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Slides were washed with 1x PBS and then blocked in 5% goat serum containing 0.3% Triton X-100. Iba-1 was stained using ab178847 at 1/500 dilution in immunohistochemical analysis. Representative immunofluorescence is depicted for KO Sham (F top left), KO injured (F bottom left), WT Sham (F top right), and WT injured (F bottom right) in the right and left hemispheres (blue = DAPI, green = Iba-1). Scale bars = 100μm, 20x. HI = hypoxic-ischemic brain injury.
Area occupied by Iba-1-positive staining was elevated in KO injured animals when compared to uninjured controls, while stain area was not elevated in WT injured animals
Iba1 was immunoprecipitated from 1mg of Mouse spleen whole cell lysate with ab178847 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using ab178847 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: Mouse spleen whole cell lysate 10μg (Input).
Lane 2: ab178847 IP in Mouse spleen whole cell lysate.
Lane 3: Rabbit IgG,monoclonal-Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab178847 in Mouse spleen whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
All lanes: Immunoprecipitation - Anti-Iba1 antibody [EPR16589] (ab178847)
Predicted band size: 16 kDa
Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized U937 (Human histiocytic lymphoma cell line) cells labeling Iba1 with ab178847 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on U937 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab178847 at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution.
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Iba1 with ab178847 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Cytoplasm staining on microglia of the normal Human cerebrum is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution.
Iba1 antibody ab178847 was used with Tissue Clearing Kit Tissue Clearing Kit - hydrophobic ab243298 to penetrate, stain and clear a 1 mm coronal section of mouse brain. Blue: DAPI , Green: Iba1.
Learn more about tissue clearing kits, reagents, and protocols designed to make it easier to stain thick tissue sections and get more data from each valuable tissue section.
For 1 mm brain sections, we recommend a starting dilution of 1:100, and also using Goat Anti-Rabbit IgG H&L AlexaFluor488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1:400.
Immunohistochemical analysis of paraffin-embedded Mouse endometrium tissue labeling Iba1 with ab178847 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Cytoplasm staining on macrophages of the mouse endometrium.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized THP-1 (Human monocytic leukemia cell line) cells labeling Iba1 with ab178847 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on THP-1 cell line.
The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab178847 at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution.
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Iba1 antibody [EPR16589] (ab178847) at 1/2000 dilution
Lane 1: Human spleen lysate at 20 µg
Lane 2: THP-1 (Human monocytic leukemia cell line) whole cell lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 16 kDa
Observed band size: 17 kDa
Exposure time: 3min
Different batches of ab178847 were tested on U-937 (Human histiocytic lymphoma monocyte) lysate at 0.5 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 17 kDa.
All lanes: Western blot - Anti-Iba1 antibody [EPR16589] (ab178847)
Predicted band size: 16 kDa
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Iba1 antibody [EPR16589] (ab178847) at 1/1000 dilution
Lane 1: MOLT-4 (Human lymphoblastic leukemia cell line) whole cell lysate at 10 µg
Lane 2: U937 (Human histiocytic lymphoma cell line) whole cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution
Predicted band size: 16 kDa
Observed band size: 17 kDa
Exposure time: 3min
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 and 2: 1 minute; Lane 3 and 4: 3 minutes.
All lanes: Western blot - Anti-Iba1 antibody [EPR16589] (ab178847) at 1/1000 dilution
Lane 1: Mouse spleen lysate at 10 µg
Lane 2: Rat spleen lysate at 10 µg
Lane 3: Mouse testis lysate at 10 µg
Lane 4: Rat testis lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution
Predicted band size: 16 kDa
Observed band size: 17 kDa
Iba1 Immunohistochemistry (Frozen sections) staining of Mouse cerebrum (perfused fixed) using rabbit Anti-Iba1 antibody
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (perfused fixed) tissue labeling Iba1 with ab178847 at 1200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/1000 dilution (Green).
Confocal image showing positive staining on mouse cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab178847 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081)at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Iba1 Western blot staining using rabbit Anti-Iba1 antibody
IBA1 is a relatively minor protein of brain and is much more abundant in spleen (PMID: 8912632, PMID: 29232670). We suggest loading higher amount of brain lysate or using lower dilution of antibody for detecting signal in brain related lysates.
All lanes: Western blot - Anti-Iba1 antibody [EPR16589] (ab178847) at 1/1000 dilution
Lane 1: Mouse spleen tissue lysate at 20 µg
Lane 2: Mouse brain tissue lysate at 20 µg
Lane 3: Mouse hippocampus tissue at 20 µg
Lane 4: Rat spleen tissue lysate at 20 µg
Lane 5: Rat brain tissue lysate at 20 µg
Lane 6: Rat hippocampus tissue lysate at 20 µg
All lanes: Western blot - PE Anti-ENPP3/B10 antibody [NP4D6] (PE Anti-ENPP3/B10 antibody [NP4D6] ab90751) at 1/20000 dilution
Predicted band size: 16 kDa
Observed band size: 17 kDa
Exposure time: 40s
Iba1 Immunohistochemistry (Frozen sections) staining of Mouse liver (perfused fixed) using rabbit Anti-Iba1 antibody
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (perfused fixed) tissue labeling Iba1 with ab178847 at 1200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/1000 dilution (Green).
Confocal image showing positive staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab178847 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Iba1 Immunohistochemistry (Frozen sections) staining of Rat cerebrum (perfused fixed) using rabbit Anti-Iba1 antibody
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum (perfused fixed) tissue labeling Iba1 with ab178847 at 1200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/1000 dilution (Green).
Confocal image showing positive staining on rat cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab178847 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
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