Overview

  • Product name

    Anti-Vitronectin/S-Protein antibody [VN58-1]
    See all Vitronectin/S-Protein primary antibodies
  • Description

    Mouse monoclonal [VN58-1] to Vitronectin/S-Protein
  • Host species

    Mouse
  • Specificity

    This antibody does not interfere with Vitronectin/S-Protein-mediated adhesion.
  • Tested applications

    Suitable for: ICC/IF, ELISA, WB, IHC-P, IHC-Frmore details
  • Species reactivity

    Reacts with: Human
    Does not react with: Cow
  • Immunogen

    Full length protein corresponding to Human Vitronectin/S-Protein.

  • Epitope

    This antibody specifically reacts with an epitope in the region of amino acids 1-130 of human Vitronectin/S-Protein.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.1% Sodium azide
    Constituents: 0.0268% PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Ascites
  • Purification notes

    Purified from ascites.
  • Clonality

    Monoclonal
  • Clone number

    VN58-1
  • Isotype

    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab13413 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 a concentration of 1 µg/ml.
ELISA 1/3000. (with solid phase antigen)
WB Use a concentration of 5 - 10 µg/ml. Predicted molecular weight: 54 kDa.
IHC-P Use a concentration of 5 - 10 µg/ml.
IHC-Fr Use a concentration of 5 - 10 µg/ml.

Target

  • Function

    Vitronectin is a cell adhesion and spreading factor found in serum and tissues. Vitronectin interact with glycosaminoglycans and proteoglycans. Is recognized by certain members of the integrin family and serves as a cell-to-substrate adhesion molecule. Inhibitor of the membrane-damaging effect of the terminal cytolytic complement pathway.
    Somatomedin-B is a growth hormone-dependent serum factor with protease-inhibiting activity.
  • Tissue specificity

    Plasma.
  • Sequence similarities

    Contains 4 hemopexin repeats.
    Contains 1 SMB (somatomedin-B) domain.
  • Domain

    The SMB domain mediates interaction with SERPINE1/PAI1. The heparin-binding domain mediates interaction with insulin.
  • Post-translational
    modifications

    Sulfated on 2 tyrosine residues.
    N- and O-glycosylated.
    Phosphorylation on Thr-69 and Thr-76 favors cell adhesion and spreading.
    It has been suggested that the active SMB domain may be permitted considerable disulfide bond heterogeneity or variability, thus two alternate disulfide patterns based on 3D structures are described with 1 disulfide bond conserved in both.
    Phosphorylation sites are present in the extracellular medium.
  • Cellular localization

    Secreted, extracellular space.
  • Information by UniProt
  • Database links

  • Alternative names

    • Complement S Protein antibody
    • Epibolin antibody
    • S Protein antibody
    • S-protein antibody
    • Serum Spreading Factor antibody
    • Serum-spreading factor antibody
    • Somatomedin B antibody
    • Somatomedin-B antibody
    • V75 antibody
    • Vitronectin antibody
    • Vitronectin V10 subunit antibody
    • Vitronectin V65 subunit antibody
    • VN antibody
    • VNT antibody
    • VTN antibody
    • VTNC_HUMAN antibody
    see all

Images

  • This picture shows formalin-fixed paraffin embedded human skin stained with Vitronectin/S-Protein (1:100 - 30 minutes RT). The image was kindly supplied as part of the review submitted by Elizabeth Chlipala.

  • ICC/IF image of ab13413 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab13413, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:

  • Simsa R  et al. Effect of fluid dynamics on decellularization efficacy and mechanical properties of blood vessels. PLoS One 14:e0220743 (2019). Read more (PubMed: 31381614) »
  • Siney EJ  et al. Metalloproteinases ADAM10 and ADAM17 Mediate Migration and Differentiation in Glioblastoma Sphere-Forming Cells. Mol Neurobiol 54:3893-3905 (2017). Read more (PubMed: 27541285) »
See all 7 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (Epithelial OVCA cell line)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
100 µg
Treatment
Perifosine 24 hr
Specification
Epithelial OVCA cell line
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Aug 12 2016

Question

Hi, i'm responding the questionary 1) Abcam product code ab: ab13413, ab27710, ab77811 2) Abcam order reference number or product batch number 3) Description of the problem: no bands 4) Sample preparation: Type of sample: HepG2 whole cell lysates Species : Human Lysis buffer : 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 5% glycerol Protease inhibitors: aprotinin, leupeptin, bestatin, pesptatin and E-64 Phosphatase inhibitors : Reducing agent : DTT Boiling for ≥5 min? yes Protein loaded ug/lane: generally 8 ug Positive control : Negative control : 5) Percentage of gel: 12% Type of membrane: Nitrocelulose Protein transfer verified: Panceau Red Blocking agent and concentration: I get no bands with non fat milk 5% + TBST, and some ghosty bands with BSA sigma 5% + TBST Blocking time: 2h for milk and overnight for BSA Blocking temperature: 4 oC 6) Primary antibody (If more than one was used, describe in “additional notes”) : Concentration or dilution generally 1:1000 Diluent buffer TBST + blocking agent Incubation time O.N. utilizing milk 5% as blocking agent and 2h utilizing BSA 5% as blocking agent. Incubation temperature: 4 oC 7) Secondary antibody: anti-mouse Species: Reacts against: Concentration or dilution , generally 1:4000 Diluent buffer TBST + blocking agent Incubation time 2h Incubation temperature: 4 oC Fluorochrome or enzyme conjugate: HRP 8) Washing after primary and secondary antibodies: Buffer TBST Number of washes 3x (10 min) 9)Detection method ECL 10) How many times have you run this staining? 3 Do you obtain the same results every time? yes What steps have you altered to try and optimize the use of this antibody? Change the blocking agent (Milk to BSA).

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Answer

Thank you for getting back to me with those details. Having reviewed your protocol there are a few things I may be able to suggest. The dilution for ab13413 is recommended in the range of 5-10µg/ml and ab27710 at an assay dependent dilution. I would therefore suggest decreasing the dilution that you are using trying 1/200 and 1/500 to see if any improvement is observed. Even though ab77811 is recommended at 1/1000 I would also suggest trying this with 1/200 and 1/500 as this may also need optimising with your sample. I would also suggest increasing the amount of protein that you are loading on the gel, we would normally recommend loading between 20-40 µg per well. As you are observing some bands with blocking using BSA I would continue to use this blocking agent. Initially using 3% in TBST incubating for 1-2 hours at room temperature. I would then incubate the primary antibody diluted in 1% BSA TBST for 1 hour at room temperature and overnight at 4°C with agitation. It may be that the conditions that you have been using has been over blocking the membrane. I would also reduce the wash step to 3x5 min. I realise this is alot to try with three different antibodies so I would recommend optimising each one at a time to find the best experimental conditions for each antibody. May I ask what secondary antibodies you have been using to detect the primary antibodies. ab13413, ab2771 and ab77811 are mouse IgG1, IgG2b and IgM respectively and would need to be used with secondary antibodies which are able to recognise these isotypes. I hope these suggestions help to improve the results you have been observing. If you continue to have problems please do let me know.

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Answer

Thank you for contacting us. I am sorry that you are experiencing problems with these antibodies. In order to assist you as best I can could you please fill in the questionnaire I have attached to this email. This will allow me to understand more clearly the experiments that you have been performing and identify any advice I can give in order to achieve better results. It would be helpful if you did this for each antibody individually and if you could provide me with any images of the blots you have obtained this would also be of help. I look forward to hearing back from you.

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