使用敲除细胞株进行验证RabMAb

Anti-TMEM119抗体[28-3] - Microglial marker (ab209064)

概述

  • 产品名称
    Anti-TMEM119抗体[28-3] - Microglial marker
    参阅全部 TMEM119 一抗
  • 描述
    兔单克隆抗体[28-3] to TMEM119 - Microglial marker
  • 特异性
    This antibody recognizes mouse Tmem119, a transmembrane protein that has been reported to be a highly specific microglia marker that is not expressed by macrophages or other immune or neural cell types (Bennett et al., 2016).
  • 经测试应用
    适用于: IHC-Fr, IHC-Pmore details
  • 种属反应性
    与反应: Mouse
    不与反应: Rat, Human
  • 免疫原

    Recombinant fragment (GST-tag) within Mouse TMEM119 aa 100 to the C-terminus (intracellular). The exact sequence is proprietary.

  • 阳性对照
    • IHC-Fr: mouse brain. IHC-P: FFPE mouse brain. Mouse brain cerebral cortex, hippocampus and cerebellum stain positive for Tmem119. Please note that Tmem119 expression is seen after postnatal day (P) 14 in mouse brain.
  • 常规说明

    This Tmem119 antibody has been knockout validated in IHC, meaning it demonstrated the expected staining in wild type mouse brain sections and no staining was observed in Tmem119 knockout mouse brain sections. The data are shown on this datasheet. To detect mouse Tmem119 by flow cytometry, we recommend using ab210405. To detect human TMEM119 by IHC, we recommend using ab185333.

    The 28-3 clone to mouse Tmem119 is exclusively manufactured and sold by Abcam.

    IHC-Frozen protocol advice:
    For immunohistochemistry on frozen sections, it is recommended that a high concentration of Triton X-100 (0.5%) is used during permeabilization and antibody incubation steps. This may increase the proportion of microglia that stain positive for Tmem119.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • 存储溶液
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Please note that some batches of ab209064 may contain 0.4M arginine. Please contact Scientific Support for further information.
  • Concentration information loading...
  • 纯度
    Protein G purified
  • 克隆
    单克隆
  • 克隆编号
    28-3
  • 同种型
    IgG

应用

Our Abpromise guarantee covers the use of ab209064 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-Fr Use a concentration of 0.5 - 1 µg/ml.

We recommend using 0.3-0.5% Triton X-100.

IHC-P Use a concentration of 0.5 - 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

靶标

  • 细胞定位
    Membrane; Single-pass type I membrane protein
  • 数据库链接
  • 别名
    • OBIF antibody
    • Osteoblast induction factor antibody
    • PSEC0199 antibody
    • Transmembrane protein 119 antibody
    • UNQ731/PRO1415 antibody
    see all

图片

  • IHC image of TMEM119 staining in a section of frozen normal mouse brain wild type (upper panel) and TMEM119 knockout (lower panel). No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in PBS containing 0.5% (v/v) Triton X-100, 0.3 M (w/v) glycine and 1% (w/v) BSA for 1 h at room temperature. The section was then incubated overnight at +4°C in PBS containing 0.5% (v/v) Triton X-100 and 1% (w/v) BSA with ab209064 at 0.5 µg/ml. The secondary antibody was ab150087 (shown in red) used at 2 µg/ml for 1 hour at room temperature. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount®. Images were taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

  • IHC image of TMEM119 and Iba1 co-staining in a section of formalin-fixed paraffin-embedded normal mouse brain. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked in PBS containing 0.5% (v/v) Triton X-100, 0.3 M (w/v) glycine and 1% (w/v) BSA for 1 h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.5% (v/v) Triton X-100 and 1% (w/v) BSA with ab209064 at 1 µg/ml and ab5076 at 5 µg/ml. The secondary antibodies were ab150087 (shown in red) and ab150133 (shown in green) used at 2 µg/ml for 1 hour at room temperature. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount®. Images were taken with a confocal microscope (Leica-Microsystems, TCS SP8).

     

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

  • ab209064 staining TMEM119 in Mouse corpus callosum sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 0.5% BSA for 1 hour at 23°C. Samples were incubated with primary antibody at 1.4µg/ml for 18 hours at 4°C. An Alexa Fluor® 488 -conjugated Donkey anti-rabbit IgG polyclonal was used as the secondary antibody.

     

    TMEM119 (green), Iba1 (red) and DAPI (blue)

    See Abreview

  • IHC image of TMEM119 staining in a section of formalin fixed, paraffin embedded normal mouse brain. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked in PBS containing 0.5% (v/v) Triton X-100, 0.3 M (w/v) glycine and 1% (w/v) BSA for 1 h at room temperature. The section was then incubated overnight at +4°C in PBS containing 0.5% (v/v) Triton X-100 and 1% (w/v) BSA with ab209064 at 0.1 µg/ml. The secondary antibody was ab150087 (shown in red) used at 2 µg/ml for 1 hour at room temperature. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount®. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

     

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

  • IHC image of TMEM119 staining in a section of frozen normal mouse brain. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in PBS containing 0.5% (v/v) Triton X-100, 0.3 M (w/v) glycine and 1% (w/v) BSA for 1 h at room temperature. The section was then incubated overnight at +4°C in PBS containing 0.5% (v/v) Triton X-100 and 1% (w/v) BSA with ab209064 at 0.5 µg/ml. The secondary antibody was ab150087 (shown in red) used at 2 µg/ml for 1 hour at room temperature. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount®. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

     

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

  • IHC image of TMEM119 staining in a section of formalin fixed, paraffin embedded normal mouse brain, 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 ab209064, 0.5 µg/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.

  • Normal (WT) mouse brain, stained for TMEM119 (red), Iba1 (green) and DAPI (blue). Samples were baked onto slides for 10 minutes at 60oC, rehydrated with PBS and blocked with blocking buffer (10% serum in PBST). ab209064 at a concentration of 1 µg/mL was incubated with the sample overnight at 4oC. Slides were washed with PBS and a goat anti-rabbit Alexa Fluor 488® was used as the secondary antibody at a concentration of 4 µg/mL.

  • Normal mouse choroid plexus, stained for TMEM119 (red), Iba1 (green) and DAPI (blue). Choroid plexus macrophages are positive for Iba1 and negative for TMEM119. Samples were baked onto slides for 10 minutes at 60oC, rehydrated with PBS and blocked with blocking buffer (10% serum in PBST). ab209064 at a concentration of 1 µg/mL was incubated with the sample overnight at 4oC. Slides were washed with PBS and a goat anti-rabbit Alexa Fluor 488® was used as the secondary antibody at a concentration of 4 µg/mL.

  • Representative FISH analysis of GAS5 (green) co‐stained with ab209064 (red) in spinal cord sections from EAE mice at 30 dpi. Arrows indicate GAS5+TMEM119+ cells. Scale bars = 25 μm.

    Female C57BL/6 mice (6–8 weeks) were deeply anesthetized with 3% chloral hydrate and a laminectomy was performed. After fixing the spine, 1 μl of 1% lysolecithin in a 0.9% sodium chloride solution was injected into the dorsal funiculus at the level of the T11–T12 vertebrae. The day of lysolecithin injection was designated day 0 (0 dpi). The spinal cord around the injection point was isolated and cut into serial cryosections.

    Tissue sections were fixed, permeabilized, and incubated with the primary antibody overnight at 4°C, followed by 2 h of incubation with TRITC‐ or FITC‐conjugated secondary antibodies. Then, the samples were counterstained with Hoechst 33342.

  • Representative images of sham (d–g) and hypoperfusion (h–k) at 12 weeks post-surgery are shown to illustrate Iba-1 immunostaining in sham (d) and hypoperfused (h); TMEM119 immunostaining in sham (e) and hypoperfused (i) and then Iba-1/TMEM119 co-localisation in sham (f,g) and hypoperfused (j,k) white matter. All Iba-1+ cells in both sham and hypoperfused cohorts were also TMEM119+ indicating that the cells in the corpus callosum were resident microglia. Scale bars; d-f and h-j are 50µm, g and k 10 µm. The number of microglial cells significantly correlated with nodal gap length.

    Free floating cryo-preserved sections cut at 30 μm thickness. Sections were incubated with the primary antibodies (anti-Iba-1 (1/100) and anti-TMEM119 (1/500, ab209064)) overnight at 4°C. Sections were stained at the outset with haematoxylin and eosin to determine the presence and absence of ischemic neuronal perikaryal damage as part of the inclusion/exclusion criteria.

文献

This product has been referenced in:
  • Manso Y  et al. Minocycline reduces microgliosis and improves subcortical white matter function in a model of cerebral vascular disease. Glia N/A:N/A (2017). IHC-Fr, IHC-FrFl ; Mouse . Read more (PubMed: 28722234) »
  • Sun D  et al. LncRNA GAS5 inhibits microglial M2 polarization and exacerbates demyelination. EMBO Rep N/A:N/A (2017). IHC ; Mouse . Read more (PubMed: 28808113) »

See all 4 Publications for this product

客户评价及客户问答

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (30um-thick brain slices)
Permeabilization
Yes - 0.3% Triton X-100
Specification
30um-thick brain slices
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative
Paraformaldehyde
Username

Francesca Mowry

Verified customer

提交于 Jan 04 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Equus Tissue sections (Cerebellum)
Permeabilization
Yes - 0.5% Triton X-100
Specification
Cerebellum
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
4% PFA for 5 mins
Username

Stephen Cahalan

Verified customer

提交于 Dec 12 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Username

Abcam user community

Verified customer

提交于 Oct 18 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (Spleen)
Gel Running Conditions
Reduced Denaturing (10%)
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
Username

Dr. Sergi Bayod

Verified customer

提交于 Sep 28 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: 23°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Mar 21 2017

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

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