製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to Iba1
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: ICC/IF, IHC-P, WB, Flow Cyt, IHC-Frmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Recombinant fragment, corresponding to a region within acids 1-147 of Human Iba1 (UniProt: P55008).

  • ポジティブ・コントロール
    • THP1, HL60 and Mouse liver whole cell lysates; primary murine microglia and HeLa cells; Human U373 xenograft tissue. Spleen tissue; U937, THP1, NR8383 whole cell lysates; Mouse and rat brain tissues. E13.5 rat brain tissue. Rat liver extract.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab153696 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 1/100 - 1/1000.
IHC-P 1/100 - 1/1000.
WB 1/500 - 1/10000. Predicted molecular weight: 16 kDa.

We recommend blocking in 3% milk. Blocking with BSA gives high background.

Flow Cyt Use at an assay dependent concentration.

Use 1 µg for 4 x 105 cells.

 

 

 

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

IHC-Fr 1/100 - 1/1000.

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

画像

  • IHC image of Iba1 staining in normal mouse brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. 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 ab153696, 1µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC 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.

  • IHC image of Iba1 staining in human normal hippocampus 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 ab153696, 5µg/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.

  • Flow cytometric analysis of THP-1 (human monocytic leukemia cell line) cell line labeling Iba1 with ab153696 at 1/50 dilution (red) compared with an unlabeled sample (blue).

  • Anti-Iba1 antibody (ab153696) at 1/1000 dilution + Rat liver extract at 50 µg

    Secondary
    HRP-conjugated anti-rabbit IgG antibody

    Predicted band size: 16 kDa

  • Frozen sectioned E13.5 Rat brain tissue stained for Iba1 using ab221591 at 1/500 dilution in immunohistochemical analysis.

  • Paraffin-embedded mouse brain tissue stained for Iba1 using ab221591 at 1/500 dilution in immunohistochemical analysis.

  • Paraffin-embedded rat brain tissue stained for Iba1 using ab221591 at 1/500 dilution in immunohistochemical analysis.

  • All lanes : Anti-Iba1 antibody (ab153696) at 1/500 dilution

    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 : 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 : MOLT4 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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 16 kDa


    Exposure time: 1 minute


    BLOCKED IN 3% MILK. For ab153696 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 ab153696 (anti-Iba1 antibody; 1/500 dilution) for 18 hours at 4°C. Antibody binding was detected using HRP-labelled anti-Rabbit IgG  for 1 hour at room temperature and visualised using ECL development solution ab133406.

  • Anti-Iba1 antibody (ab153696) at 1/1000 dilution + Mouse liver whole cell lysate at 50 µg

    Predicted band size: 16 kDa



    15% SDS PAGE
  • All lanes : Anti-Iba1 antibody (ab153696) at 1/5000 dilution

    Lane 1 : THP1 whole cell lysate
    Lane 2 : HL60 whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 16 kDa



    15% SDS PAGE
  • Flow cytometic analysis of primary murine microglia cells labeling Iba1 with ab153696 at 1.0 µg per 4×105 cells (blue), Rabbit IgG (green) , Unstained (red).
  • Immunofluorescent analysis of HeLa cells (fixed in iced cold methanol; 5mins) labeling Iba1 with ab153696 at 1/500 dilution (left). Right image shows cells co-stained with Hoechst 33342.
  • Immunohistochemical analysis of paraffin-embedded Human U373 xenograft tissue labeling Iba1 with ab153696 at 1/500 dilution.

参考文献

This product has been referenced in:
  • Tchekalarova J  et al. Chronic agomelatine treatment prevents comorbid depression in the post-status epilepticus model of acquired epilepsy through suppression of inflammatory signaling. Neurobiol Dis 115:127-144 (2018). Read more (PubMed: 29653194) »

See 1 Publication for this product

レビューと Q&A

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Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Cerebellum)
Permeabilization
Yes - 0.1% Triton X-100, 30 min
Specification
Cerebellum
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Paraformaldehyde
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投稿 Nov 18 2013

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

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