Name: Anti-Iba1 antibody [EPR16589]
Brand: RabMAb
SKU: ab178847
Rating: 4 (10 reviews)

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Primary

Secondary

This product Rabbit Anti-Iba1 antibody [EPR16589] (ab178847)
IHC-P, WB, IP, ICC/IF

Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
IHC-P, WB, ELISA, IP

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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-P, WB, IP, ICC/IFmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide within Human Iba1 aa 1-100. The exact sequence is proprietary.
Database link: P55008
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.
General notes

Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents.
This product is a recombinant rabbit monoclonal antibody.

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
- Anti-Iba1 antibody [EPR16589] (HRP) (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

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

All lanes : Anti-Iba1 antibody [EPR16589] (ab178847) at 1/200 dilution

Lane 1 : Mouse hippocampus tissue lysates
Lane 2 : Mouse brain tissue lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

Exposure time: 3 minutes

Blocking and diluting buffer: 5% NFDM/TBST.
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 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.
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.
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.
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.

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

This product has been referenced in:

Sun L et al. Differences in neurotropism and neurotoxicity among retrograde viral tracers. Mol Neurodegener 14:8 (2019). Read more (PubMed: 30736827) »
Yi SY et al. Detecting Microglial Density With Quantitative Multi-Compartment Diffusion MRI. Front Neurosci 13:81 (2019). Read more (PubMed: 30837826) »

See all 34 Publications for this product

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

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1-10 of 10 Abreviews or Q&A

Application

Immunohistochemistry (PFA perfusion fixed frozen sections)

Sample

Rat Tissue sections (Brain, hippocampus)

Antigen retrieval step

None

Permeabilization

Specification

Brain, hippocampus

Blocking step

BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: .3% · Temperature: RT°C

Fixative

Paraformaldehyde

Collin Gagne

Verified customer

Submitted Jun 21 2019

Application

Immunohistochemistry (PFA perfusion fixed frozen sections)

Sample

Mouse Tissue sections (Hippocampus)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: 10 mM Citrate buffer, pH 6.0

Permeabilization

Yes - 0.1% Triton in PBS

Specification

Hippocampus

Blocking step

1% BSA + 5% Serum + 0.3 M Glycine as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 24°C

Fixative

Paraformaldehyde

Dr. Sergi Bayod

Verified customer

Submitted Nov 19 2018

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

IHC - Wholemount

Sample

Mouse Tissue (Retinal Flat mount)

Specification

Retinal Flat mount

Mr. Tyler Kilburn

Verified customer

Submitted Mar 23 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

Application

Western blot

Sample

Mouse Tissue lysate - whole (Spleen)

Gel Running Conditions

Reduced Denaturing (4-20%)

Loading amount

20 µg

Specification

Spleen

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Dr. Sergi Bayod

Verified customer

Submitted Mar 13 2017

Application

Immunohistochemistry (Frozen sections)

Sample

Mouse Tissue sections (Hippocampus)

Permeabilization

Yes - 0.2% Triton in PBS

Specification

Hippocampus

Blocking step

1% BSA + 5% Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Acetone

Dr. Sergi Bayod

Verified customer

Submitted Mar 03 2017

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (Primary cortical neuronal cells)

Permeabilization

Yes - 0.2% Triton X-100

Specification

Primary cortical neuronal cells

Blocking step

casein solution + Tween 20 as blocking agent for 20 minute(s) · Concentration: 0.2% · Temperature: 25°C

Fixative

Ethanol

Abcam user community

Verified customer

Submitted Aug 10 2016

Application

Western blot

Sample

Human Tissue lysate - whole (Brain)

Gel Running Conditions

Reduced Denaturing (12)

Loading amount

25 µg

Specification

Brain

Blocking step

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

Abcam user community

Verified customer

Submitted Feb 11 2016

Application

Western blot

Sample

Mouse Tissue lysate - whole (Spinal Cord)

Gel Running Conditions

Reduced Denaturing (12)

Loading amount

50 µg

Treatment

MOG

Specification

Spinal Cord

Blocking step

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

Abcam user community

Verified customer

Submitted Feb 10 2016

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Recombinant Anti-Iba1 antibody [EPR16589] (ab178847)

Overview

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Datasheets and documents

References

This product has been referenced in:

Customer reviews and Q&As

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Iba1 antibody [EPR16589]

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Iba1 antibody [EPR16589]

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

IHC - Wholemount abreview for Anti-Iba1 antibody [EPR16589]

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

Western blot abreview for Anti-Iba1 antibody [EPR16589]

Immunohistochemistry (Frozen sections) abreview for Anti-Iba1 antibody [EPR16589]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Iba1 antibody [EPR16589]

Western blot abreview for Anti-Iba1 antibody [EPR16589]

Western blot abreview for Anti-Iba1 antibody [EPR16589]