製品の概要

  • 製品名Anti-Iba1 antibody [1022-5]
    Iba1 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [1022-5] to Iba1
  • 特異性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.
  • アプリケーション適用あり: ICC/IF, WB, IHC-FoFr, IHC-Pmore details
    適用なし: IHC-Fr
  • 種交差性
    交差種: Rat, Human
  • 免疫原

    Recombinant full length protein (Human).

  • ポジティブ・コントロール
    • Spleen, THP1, C6

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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-FoFr Use at an assay dependent concentration. PubMed: 20933270
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.
  • 追加情報Is unsuitable for IHC-Fr.
  • ターゲット情報

    • 機能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.
    • 組織特異性Detected in T-lymphocytes and peripheral blood mononuclear cells.
    • 配列類似性Contains 2 EF-hand domains.
    • 翻訳後修飾Phosphorylated on serine residues.
    • 細胞内局在Cytoplasm > cytoskeleton. Cell projection > ruffle membrane. Associated with the actin cytoskeleton at membrane ruffles and at sites of phagocytosis.
    • Information by UniProt
    • 参照データベース
    • 別名
      • 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 [1022-5] 画像

    • 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
      Lane 7 : 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
      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 : Rat spleen tissue lysate at 30 µg
      Lane 17 : 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 15-17: 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 Rat brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with acetone and blocked with 5% BSA for 1 hour at 37°C. The sample was incubated with primary antibody (1/100) at 4°C for 18 hours. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

      See Abreview

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

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

    Anti-Iba1 antibody [1022-5] (ab15690) 使用論文

    This product has been referenced in:
    • Liao K  et al. Cocaine-mediated induction of microglial activation involves the ER stress-TLR2 axis. J Neuroinflammation 13:33 (2016). IHC-FrFl ; Mouse . Read more (PubMed: 26860188) »
    • Rolón-Reyes K  et al. Microglia Activate Migration of Glioma Cells through a Pyk2 Intracellular Pathway. PLoS One 10:e0131059 (2015). WB ; Human . Read more (PubMed: 26098895) »

    See all 27 Publications for this product

    Product Wall

    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

    投稿 Sep 12 2013

    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

    投稿 Mar 25 2014

    This concentration is indeed quite high for an antibody so if you wish to dilute it in PBS then that is fine. I would say, that diluting it down to 1 mg/ml would be fine and is standard for most antibodies. Diluting it down to 100 ug/ml is quite a big ...

    Read More
    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

    投稿 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

    投稿 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

    投稿 Oct 18 2012

    Thank you for your enquiry.

    I would like to reassure you that our antibodies have all been tested successfully and would be covered by our guarantee in the applications listed on the datasheets. Therefore, any antibody tested in the specie...

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    Thank you for contacting us.

    Please find the information below describingour testing discount program:

    For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it ...

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    Thank you for your reply with the requested protocol information.


    I noticed there was no mention of any permeabilization or treatment with detergent. Have you tried adding Tween or Triton to TBS to perform a separate permeabilization or i...

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    Thank you for contacting Abcam regarding ab15690.


    I have reviewed the history of this antibody and we have had some reports of high background in a recent lot. These complaints have been resolved by protocol modifications or replacements ...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"