Overview

  • Product name
  • Description
    Goat polyclonal to Iba1
  • Specificity
    This antibody, ab5076, is expected to recognise the isoforms represented by NP_001614 and NP_116573 but not NP_004838 Please note: Although we have some very good Abreviews on mouse, some customers were receiving inconsistent results on mouse samples. We have therefore moved mouse to the predicted species and can no longer guarantee it. In addition, we recommend blocking in milk. Blocking with BSA gives high background. IHC-Fr. Some customers have used this product in IHC-Fr however we have had consistent complaints in this application over the last year and can no longer guarantee performance in this application. If looking for a monoclonal anti-Iba1 alternative we can recommend our RabMAb ab178846. This has the same immunogen as ab5076.
  • Tested applications
    Suitable for: Electron Microscopy, IHC-Fr, IHC-P, WB, IHC-FrFl, ICC, ICC/IFmore details
    Unsuitable for: IHC-FoFr
  • Species reactivity
    Reacts with: Rabbit, Guinea pig, Cow, Dog, Human, Pig, Common marmoset
    Predicted to work with: Mouse, Rat, Macaque monkey
  • Immunogen

    Synthetic peptide corresponding to Human Iba1 aa 135-147 (C terminal). Accession Number(s): NP_116573.1; NP_001614.3
    Sequence:

    C-TGPPAKKAISELP


    (Peptide available as ab23067)

  • General notes

    Please note: Although we have some very good Abreviews on mouse, some customers were receiving inconsistent results on mouse samples. We have therefore moved mouse to the predicted species and can no longer guarantee it.

     

    Alternative versions available:

    Anti-Iba1 antibody - C-terminal (Biotin) (ab212030)

Properties

Applications

Our Abpromise guarantee covers the use of ab5076 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
Electron Microscopy Use at an assay dependent concentration. PubMed: 26358247
IHC-Fr 1/500.
IHC-P Use a concentration of 2 - 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 0.3 - 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).Can be blocked with Human Iba1 peptide (ab23067).

We received several excellent Abreviews on this antibody working with mouse, however, mouse is not a batch tested species and we cannot guarantee that this antibody will work on mouse. Some of our customers have observed high background in mouse samples. We recommend blocking in milk. Blocking with BSA gives high background.

IHC-FrFl Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IHC-FoFr.
  • 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

    Anti-Iba1 antibody images

    • Paraffin embedded sections of human lung were stained for Iba1 with ab5076 at 5 μg/ml in immunohistochemical analysis.

    • All lanes : Anti-Iba1 antibody (ab5076) at 0.3 µg/ml

      Lane 1 : Rat Brain cell lysate (35µg protein in RIPA buffer)
      Lane 2 : Human Frontal Cortex cell lysate (35µg protein in RIPA buffer)


      Predicted band size : 17 kDa

      Primary incubation was 1 hour. Detected by chemiluminescence.

    • Free floating sections of rat striatum/microglia were stained for Iba1 with ab5076 at 1/2000 dilution in immunohistochemical analysis. Rabbit Anti-Goat IgG Biotin was used as the secondary antibody at 1/200 dilution.

    • Paraffin embedded sections of mouse brain were stained for Iba1 with ab5076 at 2 μg/ml in immunohistochemical analysis.

    • IHC-P image of Iba1 staining on rat brain sections using ab5076 (1:1000). The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 mins at 21°C. ab5076 was diluted 1:1000 and incubted with the sections for 2 hours at 21°C. The secondary antibody used was Rabbit polyclonal to anti sheep/goat IgG conjugated to biotin (1:200)

      See Abreview

    • All lanes : Anti-Iba1 antibody (ab5076) at 1 µg/ml

      Lane 1 : Human Iba1 full length recombinant protein at 0.1 µg
      Lane 2 : HEK293 whole cell lysate at 30 µg
      Lane 3 : A431 whole cell lysate at 30 µg
      Lane 4 : NIH3T3 whole cell lysate at 30 µg
      Lane 5 : Human spleen tissue lysate at 30 µg
      Lane 6 : Mouse spleen tissue lysate at 30 µg
      Lane 7 : Rat spleen tissue lysate at 30 µg
      Lane 8 : U937 whole cell lysate at 30 µg
      Lane 9 : HL60 whole cell lysate at 30 µg
      Lane 10 : THP1 whole cell lysate at 30 µg
      Lane 11 : THP1 whole cell lysate, PMA treated at 30 µg
      Lane 12 : Raw 264.7 whole cell lysate at 30 µg
      Lane 13 : C6 whole cell lysate at 30 µg
      Lane 14 : NR8383 whole cell lysate at 30 µg
      Lane 15 : Mouse spleen tissue lysate at 30 µg
      Lane 16 : THP1 whole cell lysate at 30 µg

      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 17 kDa

      Lanes 1-14: Blocked in 3% milk for 1 hour (RT). Lanes 15-16: Blocked in 5% BSA for 1 hour (RT).

      Lane 1: exposure 1 minute. Lanes 2-16: exposure 4 minutes. Abcam recommends blocking in milk for cleaner blots with reduced background, in comparison to BSA. 

      This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab5076 (anti-Iba1 antibody; 1 ug/mL) for 18 hours at 4°C. 

    • Frozen sections of mouse brain tissue were stained for Iba1 with ab5076 at 1/500 dilution in immunohistochemical analysis. Rabbit Anti-Goat IgG HRP was used as the secondary antibody at 1/500 dilution.

    • ab5076 staining Iba1 in the Macrophage cell line Mono-Mac 6 by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 20 minutes at 24°C. Samples were incubated with primary antibody (1/500 in PBS) for 16 hours at 4°C. An Alexa Fluor® 555-conjugated Rabbit anti-goat IgG polyclonal (1/1000) was used as the secondary antibody.

      See Abreview

    • IHC-P image of Iba1 staining on cat kidney sections using ab5076 (1:1000). The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 mins at 21°C. ab5076 was diluted 1:1000 and incubted with the sections for 2 hours at 21°C. The secondary antibody used was Rabbit polyclonal to anti sheep/goat IgG conjugated to biotin (1:200)

      See Abreview

    • Anti-Iba1 antibody (ab5076) at 0.3 µg/ml + Rat brain lysate at 35 µg

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

      Primary incubation was 1 hour. Detected by chemiluminescence.

    • ab5076 staining Iba1 in rat glioblastoma cell line C6 by Immunocytochemistry/ Immunofluorescence.Cells were fixed in paraformaldehyde, permeabilized using 0,1% Triton X 100 in PBS, blocked with 0.5% BSA for 30 minutes at room temperature and then incubated with ab5076 at a 1/50 dilution for 16 hours at 4°C. The secondary used was a Cy3 conjugated rabbit anti-goat polyclonal used at a 1/120 dilution. Nuclei are counterstained with DAPI.

      See Abreview

    • IHC-P image of Iba1 staining on cow kidney sections using ab5076 (1:2000). The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 mins at 21°C. ab5076 was diluted 1:2000 and incubted with the sections for 2 hours at 21°C. The secondary antibody used was Rabbit polyclonal to anti sheep/goat IgG conjugated to biotin (1:200)

      See Abreview

    • IHC-P image of Iba1 staining on marmoset bladder sections using ab5076 (1:1000). The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 mins at 21°C. ab5076 was diluted 1:1000 and incubted with the sections for 2 hours at 21°C. The secondary antibody used was Rabbit polyclonal to anti goat IgG conjugated to biotin (1:200)

      See Abreview

    • AIF1 expression in an inflammatory foci in a skin biopsy. Paraffin embedded sections were deparaffinized and boiled in 10mM citrate buffer for 30 min in a microwave to expose the AIF1 antigen, and then blocked with 5% donkey serum for 20 min. Slides were incubated for 40 min with goat-anti-AIF1 antibody (1:100) and rinsed in three changes of PBS for 1 min each.  A secondary antibody (donkey-anti-goat-FITC conjugated IgG; 1:50) was then applied to the slide for 40 min. Sections were washed again to remove the unbound antibody. The slides were counterstained with DaPI and viewed with a Nikon epi-fluorescent microscope. AIF1 positive cells appear green and the nuclei are stained blue. Numerous cells expressing the AIF1 protein are located around a vessel. (400X). Picture from Rreview by Carol Artlett submitted 30 July 2004.
    • ab5076 staining Iba1 in guinea pig brain tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in sodium citrate and then blocking with 3% serum was performed for 30 minutes at RT. The primary antibody was used at dilution at 1/200 and incubated with sample at 2% blocking serum for 18 hours at 4°C. A Biotin conjugated horse polyclonal to goat IgG was used undiluted as secondary antibody.

      See Abreview

    References for Anti-Iba1 antibody (ab5076)

    This product has been referenced in:
    • Hennessy E  et al. Systemic TNF-a produces acute cognitive dysfunction and exaggerated sickness behavior when superimposed upon progressive neurodegeneration. Brain Behav Immun 59:233-244 (2017). IHC ; Mouse . Read more (PubMed: 27633985) »
    • Arentsen T  et al. The bacterial peptidoglycan-sensing molecule Pglyrp2 modulates brain development and behavior. Mol Psychiatry 22:257-266 (2017). IHC-FoFr ; Mouse . Read more (PubMed: 27843150) »

    See all 192 Publications for this product

    Product Wall

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Rat Tissue sections (Intestine)
    Permeabilization
    Yes - 0.1% Triton X-100
    Specification
    Intestine
    Blocking step
    Cas-Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: RT°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Sep 19 2017

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Spleen)
    Permeabilization
    Yes - 0.1% Triton X-100
    Specification
    Spleen
    Blocking step
    Cas-Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: RT°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Sep 06 2017

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA PH9.0
    Permeabilization
    Yes - 0.1% TritonX-100
    Specification
    Brain
    Blocking step
    Cas-Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: RT°C
    Fixative
    Formaldehyde
    Username

    Dr. I-Neng Lee

    Verified customer

    Submitted Aug 16 2017

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Rabbit Tissue sections (PFA fixed and sucrose protected)
    Permeabilization
    No
    Specification
    PFA fixed and sucrose protected
    Blocking step
    NDS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Fixative
    Formaldehyde
    Username

    Dr. Govindaiah Vinukonda

    Verified customer

    Submitted Aug 10 2017

    Application
    Immunohistochemistry free floating
    Sample
    Rat Tissue sections (Rat striatum, microglia)
    Specification
    Rat striatum, microglia
    Username

    Dr. Edward Fletcher

    Verified customer

    Submitted Jul 13 2017

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (BRAIN)
    Permeabilization
    Yes - 0,3% TritonX
    Specification
    BRAIN
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Username

    Ewelina Palasz

    Verified customer

    Submitted Jul 11 2017

    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (Mouse brain slice)
    Antigen retrieval step
    None
    Permeabilization
    No
    Specification
    Mouse brain slice
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jun 27 2017

    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (retina)
    Antigen retrieval step
    None
    Permeabilization
    Yes - 0.3% Triton 100
    Specification
    retina
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 15°C
    Fixative
    Paraformaldehyde
    Username

    Ms. Yingdi Chen

    Verified customer

    Submitted Jun 20 2017

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Brain sections)
    Permeabilization
    Yes - 0.2 % triton X-100 in PBS
    Specification
    Brain sections
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 37°C
    Fixative
    Paraformaldehyde
    Username

    Miss. Berta Anuncibay

    Verified customer

    Submitted May 05 2017

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Horse Tissue sections (lymph node)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate pH6
    Permeabilization
    No
    Specification
    lymph node
    Blocking step
    rabbit normal serum as blocking agent for 30 minute(s) · Concentration: 15µg/mL · Temperature: 20°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Feb 02 2017

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