概述

  • 产品名称

    Anti-Iba1抗体[EPR16589]
    参阅全部 Iba1 一抗
  • 描述

    兔单克隆抗体[EPR16589] to Iba1
  • 宿主

    Rabbit
  • 经测试应用

    适用于: IHC-P, WB, IP, ICC/IFmore details
  • 种属反应性

    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide within Human Iba1 aa 1-100. The exact sequence is proprietary.
    Database link: P55008

  • 阳性对照

    • 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. IHC-Fr: Mouse hippocampus tissue.
  • 常规说明

    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 monoclonal antibody, which offers several advantages including:

    • High batch-to-batch consistency and reproducibility
    • Improved sensitivity and specificity
    • Long-term security of supply
    • Animal-free production
    For more information see here.

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能

    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.
  • 组织特异性

    Detected in T-lymphocytes and peripheral blood mononuclear cells.
  • 序列相似性

    Contains 2 EF-hand domains.
  • 翻译后修饰

    Phosphorylated on serine residues.
  • 细胞定位

    Cytoplasm > cytoskeleton. Cell projection > ruffle membrane. Associated with the actin cytoskeleton at membrane ruffles and at sites of phagocytosis.
  • Information by UniProt
  • 数据库链接

  • 别名

    • 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

图片

  • 0.1% Triton in PBS permeabilized, Paraformaldehyde-fixed frozen mouse hippocampus tissue labeling ab178847 at  1/200 dilution in immunohistochemical analysis. Goat anti-Rabbit IgG (H+L)Alexa Fluor® 488 antibody was used as the seconndary at 1/250 dilution.

    Brain fixed in 4% PFA (in PBS) at 4ºC for 24 h. Heat-induced antigen retrieval with Citrate buffer (10 mM Citric acid, 0.05% Tween 20, pH 6.0), at 90ºC for 15 min, using a water bath. Image: Iba1 ab178847 (green) and Hoechst (blue). Counterstained with Hoechst, 2 ug/ml for 5 min at room temperature. Mounted in Fluoromount-G.

  • Slides were washed with 1x PBS and then blobked in 5% goat serum containing 0.3% Triton X-100. Iba-1 was stained using ab178847 at 1/500 dilution in immunohistochemical analysis. Representative immunofluorescence is depicted for KO Sham (F top left), KO injured (F bottom left), WT Sham (F top right), and WT injured (F bottom right) in the right and left hemispheres (blue = DAPI, green = Iba-1). Scale bars = 100μm, 20x. HI = hypoxic-ischemic brain injury.

    Area occupied by Iba-1-positive staining was elevated in KO injured animals when compared to uninjured controls, while stain area was not elevated in WT injured animals 

  • 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.

  • 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 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.

  • 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 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 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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • 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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • 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.

文献

This product has been referenced in:

See all 36 Publications for this product

客户评价及客户问答

1-10 of 10 Abreviews or Q&A

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

提交于 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

提交于 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

提交于 Sep 26 2018

Application
IHC - Wholemount
Sample
Mouse Tissue (Retinal Flat mount)
Specification
Retinal Flat mount

Mr. Tyler Kilburn

Verified customer

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 Feb 10 2016

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