Anti-CD146 antibody [P1H12] (ab24577)

Reviews (8)Q&A (5)References (20)

Overview

  • Product name

    Anti-CD146 antibody [P1H12]
    See all CD146 primary antibodies

  • Description

    Mouse monoclonal [P1H12] to CD146

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, IHC-P, WB, ELISA, IHC-Fr, IP, Flow Cytmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Dog

  • Immunogen

    Tissue, cells or virus corresponding to Human CD146.

  • Positive control

    • 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.

  • General notes

    This antibody clone is manufactured by Abcam.

    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

Applications

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.

Application Abreviews Notes
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.

Target

  • Function

    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.

  • Tissue specificity

    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.

  • Sequence similarities

    Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 2 Ig-like V-type (immunoglobulin-like) domains.

  • Cellular localization

    Membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • 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

    Images

    • Flow Cytometry - Anti-CD146 antibody [P1H12] (ab24577)
      Flow Cytometry - Anti-CD146 antibody [P1H12] (ab24577)Image from Mendes LF et al., PLoS One. 2012;7(7):e41051. Epub 2012 Jul 19. Fig 1.; doi:10.1371/journal.pone.0041051; July 19, 2012, PLoS ONE 7(7): e41051.
      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).
    • Immunocytochemistry/ Immunofluorescence - Anti-CD146 antibody [P1H12] (ab24577)
      Immunocytochemistry/ Immunofluorescence - Anti-CD146 antibody [P1H12] (ab24577)Image courtesy of an anonymous 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [P1H12] (ab24577)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [P1H12] (ab24577)
      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.
    • Western blot - Anti-CD146 antibody [P1H12] (ab24577)
      Western blot - Anti-CD146 antibody [P1H12] (ab24577)
      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
      All lanes : 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.
    • Immunohistochemistry (Frozen sections) - Anti-CD146 antibody [P1H12] (ab24577)
      Immunohistochemistry (Frozen sections) - Anti-CD146 antibody [P1H12] (ab24577)This image is courtesy of an Abreview submitted by Mr Faisal Adhami

       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

    References

    This product has been referenced in:

    See all 20 Publications for this product

    Publishing research using ab24577? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-10 of 13 Abreviews or Q&A

    Western blot abreview for Anti-CD146 antibody [P1H12]

     Excellent
    Abreviews
    abreview image
    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
    Read More

    Dr. guoqing zhu

    Verified customer

    Submitted Jan 22 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD146 antibody [P1H12]

     Inconclusive
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 01 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CD146 antibody [P1H12]

     Average
    Abreviews
    abreview image
    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
    Read More

    Mark Olson

    Verified customer

    Submitted Dec 31 2014

    Western blot abreview for Anti-CD146 antibody [P1H12]

     Good
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 26 2014

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD146 antibody [P1H12]

     Good
    Abreviews
    abreview image
    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
    Read More

    Mr. Rudolf Jung

    Verified customer

    Submitted Jan 16 2014

    Question

    I hope that you are fine and well on receiving this message. I would to know the answer regarding the cross-reactivity of CD146 antibody (catalog number: AB24577). I am locating a specific subset of human MSCs within bovine tissue and thus would like to know whether the antibody solely reacts with human cells only and not bovine tissue ? Furthermore, has this been tested by yourselves ?

    I look forward to your reply and thanks for your help.

    Best regards,

    Read More
    Answer

    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 a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    Read More

    Question

    Would like to set up an ELISA to detect CD146 in human serum. CD146 might be extracellular cleaved fragment, however.

    Read More
    Answer

    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 CD146 which is most likely the form found in serum. I've attached one article describing the soluble form running around 105 kDa in SDS-PAGE, while the membrane-bound form normally runs around 120-130 kDa. Please note that there are severalglycosylation sitesso the protein might have variable molecular weight depending on glycosylation state.

    I hope this information will be useful, but please let me know if you have any questions or if there is anything else that we can do for you and I'll be happy to help.

    Read More

    Question

    Dear Sir or Madam, Thanks for your kind help. The attachment is my protocols. In your mail, you said: “Other customers have reported band of this size using this product”. Is there any data could be seen on public or provided by you? Thank you very much and best regar

    Read More
    Answer

    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: https://www.abcam.com/index.html?datasheet=75769&tab=abreviews&intabreviewid=19756 I did not receive the attachment that you have mentioned. Could you please re-send it? Please let me know if you have any questions.

    Read More

    Question

    Dear Sir or Madam, I am very grateful for your kind information providing. But I still have a problem. The attachment is the result, which shows the observed band at 72 kDa, I detected last month (the lane 1 to 4 are all tissue lysates of mouse corpus cavernosum). The result replicated for 3 times. Could you help me to analyze that how this odd result coming? Thank you very much and best regards.

    Read More
    Answer

    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 MUC18 as I have seen this protein reported at 113kDa (PMID: 21467165) and have seen other products give a band around this size. I can only assume that the larger band may be the modified form of CD146. I would like to help you with this product if I can. Would you be able to supply me with the western blotting protocol that you used? Please include lysis buffer, denaturing steps, blocking solution as well as the times, temperature of blocking and antibody incubations. Thank you. Please contact me if you have any questions. I look forward to your reply.

    Read More

    Question

    Dear Sir or Madam, I bought an Abcam product, Anti-CD146 antibody [EPR3208] (ab75769), recently. In the data sheet, “WB: 1/10000 - 1/50000. Predicted molecular weight: 72 kDa” could be read. But according to the common knowledge and another Abcam product, Anti-CD146 antibody [P1H12] (ab24577), the band of CD146 molecular should be observed at the size of approximately 113 kDa. I am quite confused about this, thus, I allowed myself to seek help from you. I would very appreciate it if you could answer my question in your busy schedule. Thank you and happy New Year.

    Read More
    Answer

    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 However, I have found as data, which will be on our datasheet soon, which shows the product giving the more accepted and expected results. I will attach this data for you. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    1-10 of 13 Abreviews or Q&A

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