Overview

  • Product name
    Anti-Vitronectin/S-Protein antibody [BV1]
    See all Vitronectin/S-Protein primary antibodies
  • Description
    Mouse monoclonal [BV1] to Vitronectin/S-Protein
  • Host species
    Mouse
  • Specificity
    The antibody binds to soluble vitronectin as well as to membrane bound vitronectin.
  • Tested applications
    Suitable for: IHC-P, WB, IP, ICC/IF, Purificationmore details
  • Species reactivity
    Reacts with: Human

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.1% Sodium azide
    Constituent: 0.2% BSA
  • Concentration information loading...
  • Purity
    Protein G purified
  • Clonality
    Monoclonal
  • Clone number
    BV1
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab7165 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-P Use at an assay dependent concentration. PubMed: 19730772
WB Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Purification Use at an assay dependent concentration.

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

References

This product has been referenced in:
  • Ahn J  et al. The metastasis gene NEDD9 product acts through integrin ß3 and Src to promote mesenchymal motility and inhibit amoeboid motility. J Cell Sci 125:1814-26 (2012). Read more (PubMed: 22328516) »
  • Knappe UJ  et al. Expression of extracellular matrix-proteins in perisellar connective tissue and dura mater. Acta Neurochir (Wien) 152:345-53 (2010). IHC-P ; Human . Read more (PubMed: 19730772) »
See all 2 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

We regret to hear about your problems with ab7165 monoclonal antibody to human vitronectin. The antibody is described in the following article: Zanetti, A et al; Clustering of vitronectin and RGD peptides on microspheres leads to engagement of integrins on the luminal aspect of endothelial cell membrane. Blood 1994, 84: 1116 The researcher of this article use human plasma vitronectin, which was purified under nondenaturing conditions as described in: Preissner, K et al; Physicochemical characterization of human S-protein and its functions in the blood coagulation system. Biochem J 1985, 231: 349. This may suggest that the concentration of vitronectin in the serum samples is too low to detect. To check this statement we would like to advise you to do a spotblot. We hope this information will be helpful to you. If you have further questions please do not hesitate to contact us.

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Answer

I have contacted the source of the antibody to find out if they can recommend a protocol. Could you please confirm you used protease inhibitors and kept the samples cold at all times during your experiment prior to boiling your samples with loading buffer? I have been informed you also have problems with detecting kininogen and I am wondering if you have enough of those two proteins in your purified samples? Do you have a positive control to test whether these antibodies work? I would recommend trying a long incubation (18h) and more concentrated antibody. I will forward to you any information I receive from the source of the antibody, my apologies for the delay. Thank you for your patience, Please let me know how you get on,

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Question

BATCH NUMBER 74939 ORDER NUMBER 58763 DESCRIPTION OF THE PROBLEM No signal or weak signal There was no signal in blots (2 seperate blots) SAMPLE Human serum, purified, 125 ug /blot PRIMARY ANTIBODY Abcam, ab7165-50, Mouse monoclonal (BV1) against human vitronectin(0.1ug/ul), Diluted in 2% milk in TBST, 1:400 and 1: 200 dilutions, incubated 1-2 hrs, washed 10 min x 3 times in TBST SECONDARY ANTIBODY Santacruz biotechnology, Goat anti mouse IgG-HRP (0.4 ug/ul), 1:20000, Diluted in 2% milk in TBST, incubated 1 hrs, washed 10 min x 3 times in TBST. DETECTION METHOD ECL, Super signal pico stable peroxide solution from PIERCE. treated for 2-3 min, Exposed to film for 2 min and and 10 min and there was no sginal on the blot. POSITIVE AND NEGATIVE CONTROLS USED I have reprobed the same blot with another antibody with similar reagents and I get good results. ANTIBODY STORAGE CONDITIONS Antibody stored at -20 degree C. Antibody was stored in the storage buffer supplied and diluted in freshly prepared 2% milk solution in TBST prior to incubation. SAMPLE PREPARATION (Serum purified by Agilent immuno affinity column, (enrcihed by depleting high abundant fractions like albumin, IgG, haptoglobin, transferin etc) AMOUNT OF PROTEIN LOADED 125 ug / blot ELECTROPHORESIS/GEL CONDITIONS SDS PAGE, Tris glycine buffer, reducing gel, 8-16% TRANSFER AND BLOCKING CONDITIONS Wet transfer using Biorad Trans blot apparatus, 4 hrs at 55 V, Tris glycine with 10% methanol, blocked in 5% milk HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? I tried 2 different experiment with 2 blots and 2 dilutions. I do similar methods with several other antibodies from abcam and other vendors and they work OK ADDITIONAL NOTES I have repeated 2 times with two different blots. I used 0.5 ug/ ml and 1 ug/ml dilution of the primary antibodies. It looks like the primary antibody is not at all binding. I have done parallel experiment with other antibodies and work well. Let me know ASAP what else I can do on this. Thanking you

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Answer

I'm sorry you are having problems with ab7165 and would like to thank you for taking the time to give me details of your protocol, it is very useful. I would like to suggest incubating overnight at 4C and try also more concentrated antibody. Could it be possible that you have low levels of vitronectin in your samples which cannot be detected by western blot methods? Do you have a positive control to help determine this? Do you use your secondary antibody with other antibodies? If not, could this be the problem? Degradation of vitronectin could also be the problem. Could it be degraded during the purification process and do you have protease inhibitors in your lysis buffer? Please let me know if this advice helps and do not hesitate to contact us for further information,

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Answer

The subunit recognise by this antibody is undetermined, as is it's cross-reactivity with mouse.

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

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