Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-TMEM119 antibody [28-3] - BSA and Azide free (ab234501)

Overview

  • Product name

    Anti-TMEM119 antibody [28-3] - BSA and Azide free
    See all TMEM119 primary antibodies
  • Description

    Rabbit monoclonal [28-3] to TMEM119 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-Fr, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Rat, Human
  • Immunogen

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

  • Positive control

    • 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.
  • General notes

    ab234501 is the carrier-free version of ab209064 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab234501 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot. Store at +4°C. Do Not Freeze.
  • Storage buffer

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    28-3
  • Isotype

    IgG

Applications

Our Abpromise guarantee covers the use of ab234501 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
IHC-Fr Use at an assay dependent concentration.

We recommend using 0.3-0.5% Triton X-100. Perform heat mediated antigen retrieval before IHC-Fr staining protocol, if the signal is too weak.

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Tris-EDTA buffer preferred.

Target

  • Cellular localization

    Membrane; Single-pass type I membrane protein
  • Database links

  • Alternative names

    • OBIF antibody
    • Osteoblast induction factor antibody
    • PSEC0199 antibody
    • Transmembrane protein 119 antibody
    • UNQ731/PRO1415 antibody
    see all

Images

  • ab209064 at 1:2000 staining TMEM119 antibody in mouse cerebrum tissue by immunohistochemistry (FFPE). Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling TMEM119 with ab209064 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP). Positive staining on glial cells in mouse cerebrum is observed. Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0) before commencing with IHC staining protocol. Counter stained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209064).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209064).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209064).

  • 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)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209064).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209064).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209064).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209064).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209064).

  • This IHC data was generated using the same anti-TMEM119 antibody clone, 28-3, in a different buffer formulation (cat# ab209064).

    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.

  • This IHC data was generated using the same anti-TMEM119 antibody clone, 28-3, in a different buffer formulation (cat# ab209064).

    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.

  • This IHC data was generated using the same anti-TMEM119 antibody clone, 28-3, in a different buffer formulation (cat# ab209064).

    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.

References

ab234501 has not yet been referenced specifically in any publications.

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