Name: Anti-Iba1 antibody [EPR16589]
Brand: RabMAb
SKU: ab178847
Rating: 4 (12 reviews)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] (ab178847)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16589] to Iba1
Suitable for: IHC (PFA fixed), IHC-P, WB, IP, ICC/IF
Reacts with: Mouse, Rat, Human

Product name

Anti-Iba1 antibody [EPR16589]
See all Iba1 primary antibodies
Description

Rabbit monoclonal [EPR16589] to Iba1
Host species

Rabbit
Tested applications

Suitable for: IHC (PFA fixed), IHC-P, WB, IP, ICC/IFmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human spleen lysate; THP-1, MOLT-4 and U937 whole cell lysates; Mouse and rat spleen and testis lysates; Mouse hippocampus and brain lysates. IHC-P: Human cerebrum, mouse endometrium and rat cerebrum tissues. ICC/IF: U937 and THP-1 cells. IP: Mouse spleen whole cell lysate. IHC-Fr: Mouse hippocampus tissue.
General notes
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16589
Isotype

IgG
Research areas

Alternative Versions
- HRP Anti-Iba1 antibody [EPR16589] (ab216544)
- Anti-Iba1 antibody [EPR16589] - BSA and Azide free (ab221790)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human Iba1 protein (ab105593)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab178847 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
IHC (PFA fixed)		Use at an assay dependent concentration.
IHC-P	(4)	1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB	(3)	1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).
IP		1/40.
ICC/IF	(1)	1/100.

Notes
IHC (PFA fixed) Use at an assay dependent concentration.
IHC-P 1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).
IP 1/40.
ICC/IF 1/100.

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.
Tissue specificity

Detected in T-lymphocytes and peripheral blood mononuclear cells.
Sequence similarities

Contains 2 EF-hand domains.
Post-translational
modifications

Phosphorylated on serine residues.
Cellular localization

Cytoplasm > cytoskeleton. Cell projection > ruffle membrane. Associated with the actin cytoskeleton at membrane ruffles and at sites of phagocytosis.
Target information above from: UniProt accession P55008 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 199 Human
- Entrez Gene: 11629 Mouse
- Entrez Gene: 29427 Rat
- Omim: 601833 Human
- SwissProt: P55008 Human
- SwissProt: O70200 Mouse
- SwissProt: P55009 Rat
- Unigene: 76364 Human
- Unigene: 10747 Mouse
- Unigene: 32080 Rat
see all
Alternative names
- AIF 1 antibody
- AIF-1 antibody
- Aif1 antibody
- AIF1 protein antibody
- AIF1_HUMAN antibody
- Allograft inflammatory factor 1 antibody
- Allograft inflammatory factor 1 splice variant G antibody
- allograft inflammatory factor-1 splice variant Hara-1 antibody
- balloon angioplasty responsive transcription antibody
- BART 1 antibody
- G1 antibody
- G1 putative splice variant of allograft inflamatory factor 1 antibody
- IBA 1 antibody
- IBA1 antibody
- interferon gamma responsive transcript antibody
- Interferon responsive transcript 1 antibody
- interferon responsive transcript factor 1 antibody
- Ionized calcium binding adapter molecule 1 antibody
- Ionized calcium-binding adapter molecule 1 antibody
- ionized calcium-binding adapter molecule antibody
- IRT 1 antibody
- IRT1 antibody
- Microglia response factor antibody
- MRF1 antibody
- Protein g1 antibody
see all

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] (ab178847)Stephanie R. Beldick et al.,PLOS ONE.,Fig 5.; doi.org/10.1371/journal.pone.0208105.

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
Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody [EPR16589] (ab178847)

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) (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 (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows:-

-ve control 1: ab178847 at 1/100 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Immunoprecipitation - Anti-Iba1 antibody [EPR16589] (ab178847)

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) (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 (ab172730) instead of ab178847 in Mouse spleen whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 5 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] (ab178847)

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) (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 ab97051 at 1/500 dilution.
Immunohistochemistry (PFA fixed) - Anti-Iba1 antibody [EPR16589] (ab178847)

Iba1 antibody ab178847 was used with Tissue Clearing Kit 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 (ab150077) at a dilution of 1:400.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] (ab178847)

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) (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) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody [EPR16589] (ab178847)

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) (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 ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and 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 ab150120 at 1/1000 dilution.

-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Western blot - Anti-Iba1 antibody [EPR16589] (ab178847)

All lanes : Anti-Iba1 antibody [EPR16589] (ab178847) at 1/2000 dilution

Lane 1 : Human spleen lysate
Lane 2 : THP-1 (Human monocytic leukemia cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 17 kDa
Observed band size: 17 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Iba1 antibody [EPR16589] (ab178847)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] (ab178847)

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) (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) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Iba1 antibody [EPR16589] (ab178847)

All lanes : Anti-Iba1 antibody [EPR16589] (ab178847) at 1/1000 dilution

Lane 1 : MOLT-4 (Human lymphoblastic leukemia cell line) whole cell lysate
Lane 2 : U937 (Human histiocytic lymphoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 17 kDa
Observed band size: 17 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Iba1 antibody [EPR16589] (ab178847)

All lanes : Anti-Iba1 antibody [EPR16589] (ab178847) at 1/1000 dilution

Lane 1 : Mouse spleen lysate
Lane 2 : Rat spleen lysate
Lane 3 : Mouse testis lysate
Lane 4 : Rat testis lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 17 kDa
Observed band size: 17 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1 and 2: 1 minute; Lane 3 and 4: 3 minutes.
Anti-Iba1 antibody [EPR16589] (ab178847)

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab178847? Please let us know so that we can cite the reference in this datasheet.

ab178847 has been referenced in 85 publications.

Parikh D et al. Allograft inflammatory factor-1-like is not essential for age dependent weight gain or HFD-induced obesity and glucose insensitivity. Sci Rep 10:3594 (2020). PubMed: 32107417
Yue D et al. MicroRNA-7, synergizes with RORa, negatively controls the pathology of brain tissue inflammation. J Neuroinflammation 17:28 (2020). PubMed: 31959187
Fan H et al. MsrA Suppresses Inflammatory Activation of Microglia and Oxidative Stress to Prevent Demyelination via Inhibition of the NOX2-MAPKs/NF-?B Signaling Pathway. Drug Des Devel Ther 14:1377-1389 (2020). PubMed: 32308370
Clé M et al. Study of Usutu virus neuropathogenicity in mice and human cellular models. PLoS Negl Trop Dis 14:e0008223 (2020). PubMed: 32324736
Wu D et al. Interleukin-18 from neurons and microglia mediates depressive behaviors in mice with post-stroke depression. Brain Behav Immun N/A:N/A (2020). PubMed: 32272223

View all Publications for this product

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1-4 of 4 Abreviews or Q&A

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Rat Tissue sections (Brain)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH9

Permeabilization

Specification

Brain

Blocking step

Serum as blocking agent for 20 minute(s) · Concentration: 2.5% · Temperature: 23°C

Fixative

Formaldehyde

Mr. Victor Bernal

Verified customer

Submitted Nov 09 2020

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (Brain)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate pH6

Permeabilization

Specification

Brain

Blocking step

(agent) for 30 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 31 2020

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (Brain)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Epitope retrieval 2 solution (pH=9)

Permeabilization

Specification

Brain

Fixative

Paraformaldehyde

Zohar Gavish

Verified customer

Submitted Sep 26 2018

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (Mouse brain section)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris-EDTA (pH9)

Permeabilization

Yes - 0.1% tween20

Specification

Mouse brain section

Blocking step

5% BSA and 10% serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Mar 06 2018

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (85)

Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Iba1 antibody [EPR16589]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Iba1 antibody [EPR16589]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Iba1 antibody [EPR16589]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Iba1 antibody [EPR16589]

Post-translational
modifications