Name: Anti-Iba1 antibody [EPR16589] - BSA and Azide free
SKU: ab221790
Rating: 3 (1 reviews)

Anti-Iba1 [EPR16589] antibody (ab221790) is a rabbit monoclonal antibody that is provided in a PBS only formulation free from BSA and azide. Formulated to be conjugation-ready. It is used to detect Iba1 in Western Blot, IP, IHC-P, IHC-Fr, ICC/IF. Suitable for Human, Mouse, Rat samples.

- BSA,?sodium azide, and glycerol-free?for consistent conjugation with fluorochromes, enzymes and more

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Images

Immunoprecipitation - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (AB221790), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	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

Tested
Tested

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat, Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat, Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information AIF1

Function

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.

Alternative names

G1, IBA1, AIF1, Allograft inflammatory factor 1, AIF-1, Ionized calcium-binding adapter molecule 1, Protein G1

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR16589
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

Find all reagents to label astrocytes in our "Microglia markers guide".

ab221790 is the carrier-free version of Anti-Iba1 antibody [EPR16589] ab178847.

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:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Biological function summary

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.

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

12 product images

Immunoprecipitation - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
Iba1 was immunoprecipitated from 1mg of Mouse spleen whole cell lysate with Anti-Iba1 antibody [EPR16589] ab178847 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using Anti-Iba1 antibody [EPR16589] 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: Anti-Iba1 antibody [EPR16589] ab178847 IP in Mouse spleen whole cell lysate.
Lane 3: Rabbit IgG,monoclonal-Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Iba1 antibody [EPR16589] ab178847 in Mouse spleen whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Iba1 antibody [EPR16589] ab178847).
All lanes: Immunoprecipitation - Anti-Iba1 antibody [EPR16589] (Anti-Iba1 antibody [EPR16589] ab178847)
Predicted band size: 16 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized U937 (Human histiocytic lymphoma cell line) cells labeling Iba1 with Anti-Iba1 antibody [EPR16589] 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: Anti-Iba1 antibody [EPR16589] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Iba1 antibody [EPR16589] ab178847).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Iba1 with Anti-Iba1 antibody [EPR16589] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Iba1 antibody [EPR16589] ab178847).
Immunohistochemistry (PFA fixed) - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
This data was developed using Anti-Iba1 antibody [EPR16589] ab178847, the same antibody clone in a different buffer formulation.
Iba1 antibody Anti-Iba1 antibody [EPR16589] 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
Immunohistochemical analysis of paraffin-embedded Mouse endometrium tissue labeling Iba1 with Anti-Iba1 antibody [EPR16589] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Iba1 antibody [EPR16589] ab178847).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized THP-1 (Human monocytic leukemia cell line) cells labeling Iba1 with Anti-Iba1 antibody [EPR16589] 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: Anti-Iba1 antibody [EPR16589] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Iba1 antibody [EPR16589] ab178847).
Western blot - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
This data was developed using Anti-Iba1 antibody [EPR16589] ab178847, the same antibody clone in a different buffer formulation. Different batches of Anti-Iba1 antibody [EPR16589] 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] (Anti-Iba1 antibody [EPR16589] ab178847)
Predicted band size: 16 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Iba1 with Anti-Iba1 antibody [EPR16589] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Iba1 antibody [EPR16589] ab178847).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Frozen sections) - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
Iba1 Immunohistochemistry (Frozen sections) staining of Rat cerebrum (perfused fixed) using rabbit Anti-Iba1 antibody
This data was developed using Anti-Iba1 antibody [EPR16589] ab178847, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum (perfused fixed) tissue labeling Iba1 with Anti-Iba1 antibody [EPR16589] 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 Anti-Iba1 antibody [EPR16589] 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).
Western blot - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
Iba1 Western blot staining using rabbit Anti-Iba1 antibody
This data was developed using Anti-Iba1 antibody [EPR16589] ab178847, the same antibody clone in a different buffer formulation.
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] (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
Secondary
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
Immunohistochemistry (Frozen sections) - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
Iba1 Immunohistochemistry (Frozen sections) staining of Mouse cerebrum (perfused fixed) using rabbit Anti-Iba1 antibody
This data was developed using Anti-Iba1 antibody [EPR16589] ab178847, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (perfused fixed) tissue labeling Iba1 with Anti-Iba1 antibody [EPR16589] 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 Anti-Iba1 antibody [EPR16589] 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).
Immunohistochemistry (Frozen sections) - Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
Iba1 Immunohistochemistry (Frozen sections) staining of Mouse liver (perfused fixed) using rabbit Anti-Iba1 antibody
This data was developed using Anti-Iba1 antibody [EPR16589] ab178847, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (perfused fixed) tissue labeling Iba1 with Anti-Iba1 antibody [EPR16589] 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 Anti-Iba1 antibody [EPR16589] 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).