Overview

  • Product name
    Anti-Iba1 antibody [1022-5]
    See all Iba1 primary antibodies
  • Description
    Mouse monoclonal [1022-5] to Iba1
  • Host species
    Mouse
  • Specificity
    Some customers have reported successful staining using this antibody on frozen sections, however ab15690 is not routinely tested on frozen sections and we do not recommend this antibody for use with frozen tissue. In our hands this product worked well in WB under standard reducing and denaturing conditions (please see image below). ab15690 detected Iba1 migrating at approximately 17 kDa in positive control samples (Rat spleen, THP1, C6) and bound no protein in samples we believe have little or no Iba1 expression (HEK293, NIH3T3 and A431). We recommend blocking in milk. Blocking with BSA gives high background.
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Pmore details
    Unsuitable for: IHC-Fr
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Recombinant full length protein (Human).

  • Positive control
    • Spleen, THP1, C6

Properties

Applications

Our Abpromise guarantee covers the use of ab15690 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
ICC/IF Use at an assay dependent concentration. PubMed: 24916922
WB Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).

We recommend blocking in milk. Blocking with BSA gives high background.

IHC-P Use a concentration of 10 - 20 µg/ml. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for IHC-Fr.
  • 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

    • IHC image of Iba1 staining in human normal spleen formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab15690, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

       

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • All lanes : Anti-Iba1 antibody [1022-5] (ab15690) at 1 µg/ml

      Lane 1 : Human Iba1 full length recombinant protein at 0.1 µg
      Lane 2 : HEK293 whole cell lysate at 20 µg
      Lane 3 : A431 whole cell lysate at 20 µg
      Lane 4 : NIH3T3 whole cell lysate at 30 µg
      Lane 5 : Human spleen tissue lysate at 20 µg
      Lane 6 : Mousespleen tissue lysate at 30 µg
      Lanes 7 & 16 : Rat spleen tissue lysate at 30 µg
      Lane 8 : U937 whole cell lysate at 20 µg
      Lane 9 : MOLT4 whole cell lysate at 20 µg
      Lanes 10 & 17 : 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

      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 15-17: Blocked in 5% BSA for 1 hour (RT).

      Lane 1: exposure 1 minute. Lanes 2-17: exposure 8 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 ab15690 (anti-Iba1 antibody; 1 ug/mL) for 18 hours at 4°C. Antibody binding was detected using ab97040 (HRP-labelled goat anti-mouse IgG) at  1:50,000 dilution for 1 hour at room temperature and visualised using ECL development solution ab133406.

    • Ab15690 staining IBA1 in human induced microglia by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with 4% paraformaldehyde for 15 minutes. Samples were incubated with primary antibody (1:200) for 2 hours at room temperature. Hoechst nuclear stain (blue) was used at 1:5000 dilution. 

    • Immunohistochemical analysis of paraffin embedded human speen sections, labelling Iba1 with ab15690.

    • Immunohistochemical analysis of paraffin embedded human speen sections, labelling Iba1 with ab15690.

    References

    This product has been referenced in:
    • Maiti P  et al. Solid lipid curcumin particles provide greater anti-amyloid, anti-inflammatory and neuroprotective effects than curcumin in the 5xFAD mouse model of Alzheimer's disease. BMC Neurosci 19:7 (2018). Read more (PubMed: 29471781) »
    • Ding HG  et al. Hypercapnia induces IL-1ß overproduction via activation of NLRP3 inflammasome: implication in cognitive impairment in hypoxemic adult rats. J Neuroinflammation 15:4 (2018). Read more (PubMed: 29304864) »

    See all 48 Publications for this product

    Customer reviews and Q&As

    Filter by Application

    Filter by Species

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    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (brain sections)
    Specification
    brain sections
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5 · Temperature: 37°C
    Fixative
    Acetone
    Username

    Abcam user community

    Verified customer

    Submitted Sep 12 2013

    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (Spinal cord)
    Antigen retrieval step
    None
    Permeabilization
    Yes - PBS + 0.2% triton + 4% BSA
    Specification
    Spinal cord
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Username

    Dr. Alexandre Magno

    Verified customer

    Submitted Jul 20 2018

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rhesus monkey Tissue sections (retina)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate pH 6
    Permeabilization
    No
    Specification
    retina
    Blocking step
    Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Mar 26 2018

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Sample
    Pig Cell (Brain)
    Specification
    Brain
    Permeabilization
    Yes - Sodium Citrate Buffer (10mM Sodium Citrate, 0.05% Tween 20, pH 6.0) and a pressure cooker Temp 91-99C.
    Fixative
    Formaldehyde
    Username

    Shadi Malaeb

    Verified customer

    Submitted Mar 25 2014

    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (15%)
    Sample
    Rat Cell lysate - whole cell (cortical)
    Specification
    cortical
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 07 2013

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rabbit Tissue sections (Brain)
    Specification
    Brain
    Fixative
    10% NBF
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
    Permeabilization
    Yes - Proteinase K
    Blocking step
    Serum as blocking agent for 5 minute(s) · Concentration: 0.05% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Feb 08 2013

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Brain-microglia)
    Gel Running Conditions
    Reduced Denaturing (12% gel)
    Loading amount
    35 µg
    Specification
    Brain-microglia
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 18 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (jejunum)
    Specification
    jejunum
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citric acid pH6
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
    Username

    Mr. Carl Hobbs

    Verified customer

    Submitted Apr 18 2008

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (brain)
    Specification
    brain
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid pH6
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
    Username

    Mr. Carl Hobbs

    Verified customer

    Submitted Apr 18 2008

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citric acid pH6
    Permeabilization
    No
    Specification
    brain
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
    Fixative
    Formaldehyde
    Username

    Mr. Carl Hobbs

    Verified customer

    Submitted Apr 17 2008

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