製品の概要

  • 製品名Anti-CD146 antibody [P1H12]
    CD146 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [P1H12] to CD146
  • アプリケーション適用あり: ICC/IF, IHC-P, WB, ELISA, IHC-Fr, IP, Flow Cytmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse, Dog
  • 免疫原

    Tissue/ cell preparation: human umbilical vein endothelial cells (HUVECs).

  • ポジティブ・コントロール
    • In Western Blot, ab24577 gave a positive signal in HUVEC whole cell lysate and human artery membrane lysate. In IHC, this antibody gave a positive signal in formalin-fixed paraffin-embedded human aorta tissue sections.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab24577 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.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 72 kDa).
ELISA Use a concentration of 1 - 10 µg/ml.
IHC-Fr 1/1000. See Abreview.
IP 1/150.
Flow Cyt 1/1000.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能Plays a role in cell adhesion, and in cohesion of the endothelial monolayer at intercellular junctions in vascular tissue. Its expression may allow melanoma cells to interact with cellular elements of the vascular system, thereby enhancing hematogeneous tumor spread. Could be an adhesion molecule active in neural crest cells during embryonic development. Acts as surface receptor that triggers tyrosine phosphorylation of FYN and PTK2, and a transient increase in the intracellular calcium concentration.
  • 組織特異性Detected in endothelial cells in vascular tissue throughout the body. May appear at the surface of neural crest cells during their embryonic migration. Appears to be limited to vascular smooth muscle in normal adult tissues. Associated with tumor progression and the development of metastasis in human malignant melanoma. Expressed most strongly on metastatic lesions and advanced primary tumors and is only rarely detected in benign melanocytic nevi and thin primary melanomas with a low probability of metastasis.
  • 配列類似性Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 2 Ig-like V-type (immunoglobulin-like) domains.
  • 細胞内局在Membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • A32 antigen antibody
    • CD 146 antibody
    • CD146 antibody
    • CD146 antigen antibody
    • Cell surface glycoprotein MUC18 antibody
    • Cell surface glycoprotein P1H12 antibody
    • Gicerin antibody
    • Mcam antibody
    • Melanoma adhesion molecule antibody
    • Melanoma associated antigen A32 antibody
    • Melanoma associated antigen MUC18 antibody
    • Melanoma associated glycoprotein MUC18 antibody
    • Melanoma cell adhesion molecule antibody
    • Melanoma-associated antigen A32 antibody
    • Melanoma-associated antigen MUC18 antibody
    • MelCAM antibody
    • MUC 18 antibody
    • MUC18 antibody
    • MUC18_HUMAN antibody
    • S endo 1 antibody
    • S endo 1 endothelial associated antigen antibody
    • S-endo 1 endothelial-associated antigen antibody
    see all

Anti-CD146 antibody [P1H12] 画像

  • IHC image of CD146 staining in human aorta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab24577, 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.
  • All lanes : Anti-CD146 antibody [P1H12] (ab24577) at 5 µg/ml

    Lane 1 : HUVEC (Human Umbilical Vein Endothelial Cell) Whole Cell Lysate
    Lane 2 : Human blood vessel: artery normal tissue lysate - membrane extract (ab28989)

    Lysates/proteins at 25 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 72 kDa
    Observed band size : 110 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 55 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes
    This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab24577 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
  •  ab24577 at 1/1000 dilution staining mouse brain tissue sections by Immunohistochemistry (Frozen sections). Mice were processed by transcardial perfusion first with saline, then 4% PF. After overnight incubation in 4% PF, brains were transfered to sucrose. Upon saturation, brains were frozen, sectioned with a cryostat, and then the sections were immediately mounted on slides. The tissue was incubated with ab24577 for 2 hours and then an Alexa Fluor ® 594 goat anti-mouse IgG was used as the secondary (red). DAPI staining is shown in blue. Images were taken with a confocal microscope in comparable cortex regions of the lesion or contralateral side in the same section. The lesion image shown is from this model of ischemia-hypoxia, with 1 hour of recovery time after injury, when endothelial cell activation is quite robust.

    See Abreview

  • ab24577 staining CD146 in murine bone marrow leukocytes by Immunocytochemistry/ Immunofluorescence. The cells were fixed in methanol and then blocked using 5% serum for 2 hours at 25°C. Samples were then incubated with the primary antibody at 1/400 for 12 hours at 4°C. The secondary antibody used was a goat anti-mouse IgG conjugated to Alexa Fluor® 594 (red) used at a 1/500 dilution.

    See Abreview

  • Flow cytometry analysis of hBMSCs cultured in complete a-MEM supplemented with 1 ng/mL TGF-ß1 for 7 days, staining CD146 with ab24577.

    Cells were trypsinized, counted and resuspended in 2% BSA at a concentration of 2500 cells/µL. Cells were first incubated for 45 min at 4°C, protected from light, with primary antibody antibody (1/100). Cells were then incubated for 45 minutes, protected from light, at room temperature with AlexaFluor®488-conjugated goat anti-mouse secondary antibody (1/500).

Anti-CD146 antibody [P1H12] (ab24577) 使用論文

This product has been referenced in:
  • Toshner M  et al. Transcript analysis reveals a specific HOX signature associated with positional identity of human endothelial cells. PLoS One 9:e91334 (2014). IHC-P ; Human . Read more (PubMed: 24651450) »
  • Chen SY  et al. Isolation and characterization of mesenchymal progenitor cells from human orbital adipose tissue. Invest Ophthalmol Vis Sci 55:4842-52 (2014). Read more (PubMed: 24994870) »

See all 8 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (BEL 7404)
Gel Running Conditions Non-reduced Denaturing (10%)
Loading amount 40 µg
Treatment 80 µM Doxorubicin for 24hrs
Specification BEL 7404
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. guoqing zhu

Verified customer

投稿 Jan 22 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (subcutaneous amelanotic melanoma)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: sodium citrate buffer pH6.0
Permeabilization Yes - TBS with 0.1% Tween
Specification subcutaneous amelanotic melanoma
Blocking step Milk as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Jul 01 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Sample Baboon Cell (Placental Villus)
Specification Placental Villus
Permeabilization Yes - Triton X-100 (0.1%)
Fixative 10% Buffered Formalin
Username

Mark Olson

Verified customer

投稿 Dec 31 2014

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (Hep G2)
Specification Hep G2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

投稿 Dec 26 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: TRIS-EDTA- Buffer pH 8,5
Sample Human Tissue sections (Skin)
Specification Skin
Permeabilization Yes - Washbuffer from Dako with Tween
Fixative Paraformaldehyde
Username

Mr. Rudolf Jung

Verified customer

投稿 Jan 16 2014

Thank you for contacting us.

In order to recommend the right antibody could you send us the protein sequence of Bovine CD146?

I will look forward to hearing from you soon.

Free Rabbit monoclonal antibody with any purchase of ...

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Thank you for your call yesterday and for your patience while I have looked into your enquiry.

I haven't found anything specific regarding the molecular weight of CD146 in serum, but I have found some literature regarding a soluble form of CD...

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Thank you for your response. We do have an Abreview on our website from a customer who also reported a band of that approximate size. The link is: http://www.abcam.com/index.html?datasheet=75769&tab=abreviews&intabreviewid=19756 I did not receive th...

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Thank you for contact Abcam. The expected size for this target, the L isoform, is 72kDa (Swiss Prot: http://www.uniprot.org/uniprot/Q8R2Y2). Other customers have reported band of this size using this product. There seems to be some confusion as to the ...

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Thank you for contacting us. I believe that the information presented on the datasheet if from the predicted molecular weight information available at Swiss Prot. I've attached a link to that data below: http://www.uniprot.org/uniprot/P43121#P43121 Ho...

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