Overview

  • 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.

Properties

Applications

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.

Target

  • 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.
  • Information by UniProt
  • Database links
  • 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

Images

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

References

This product has been referenced in:
See all 34 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

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1-10 of 10 Abreviews

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain, hippocampus)
Antigen retrieval step
None
Permeabilization
No
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
No
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

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