|Application:||IHC-P, IHC-F, WB|
Data & Images
|Product Name||Anti-VWF Picoband™ Antibody|
|Description||Rabbit IgG polyclonal antibody for von Willebrand factor(VWF) detection. Tested with WB, IHC-P, IHC-F in Human;Mouse.|
|Cite This Product||Anti-VWF Picoband™ Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9062)|
|Replacement Item||This antibody may replace the following items: sc-59810|sc-14014|sc-65973|sc-271409|sc-53466|sc-53465|sc-8068|sc-365712|sc-59957|sc-21784|sc-73268 from Santa Cruz Biotechnology.|
|Validated Species||Human, Mouse|
|Application||IHC-P, IHC-F, WB
*Our Boster Guarantee covers the use of this product in the above tested applications.
**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.
|Recommended Detection Systems||Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P) and IHC(F).
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2017!
|Immunogen||E.coli-derived human VWF recombinant protein (Position: R2535-K2813). Human VWF shares 79% amino acid (aa) sequence identity with mouse VWF.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
*carrier free antibody available upon request.
|Concentration||Add 0.2ml of distilled water will yield a concentration of 500ug/ml.|
|Storage||At -20˚C for one year. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for a longer time.Avoid repeated freezing and thawing.|
|Purification||Immunogen affinity purified.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Protein Name||von Willebrand factor|
|Molecular Weight||309265 MW|
|Protein Function||Important in the maintenance of hemostasis, it promotes adhesion of platelets to the sites of vascular injury by forming a molecular bridge between sub-endothelial collagen matrix and platelet-surface receptor complex GPIb-IX-V. Also acts as a chaperone for coagulation factor VIII, delivering it to the site of injury, stabilizing its heterodimeric structure and protecting it from premature clearance from plasma.|
|Sequence Similarities||Contains 1 CTCK (C-terminal cystine knot-like) domain.|
|Subcellular Localization||Secreted . Secreted, extracellular space, extracellular matrix . Localized to storage granules.|
|Alternative Names||von Willebrand factor;vWF;von Willebrand antigen 2;von Willebrand antigen II;VWF;F8VWF;|
|Research Areas|||cardiovascular|blood|serum proteins| cardiovascular|platelets|coagulation|intrinsic|extrinsic|regulatory| cancer|invasion/microenvironment|angiogenesis|angiogenic growth factors| stem cells|endothelial progenitors|endothelial markers| kits/ lysates/ other|elisa kits|blood coagulation elisa kits|cardiovascular elisa kits||
Background for von Willebrand factor
Dilution Ratios/Recommended Concentrations
At Boster we strive to provide the best Anti-VWF Picoband™ Antibody by testing all applications on non-spiked tissues and cell lines to ensure that the affinity of the antibody is enough to react to the endogenouse level of the target protein. Read more about our QC panel here.
|Recommended dilution ratios are listed below:|
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, By Heat|
Immunohistochemistry(Frozen Section), 0.5-1μg/ml
Western blot, 0.1-0.5μg/ml
**Boster provides high sensitivity secondary antibody kits for Western blotting and IHC. For more info see Related Products below.
Anti-VWF Picoband™ Antibody Images
Click the images to enlarge.
VWF was detected in paraffin-embedded section of Human Lung Cancer Tissue . Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-VWF Antibody (PB9062) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: HT1080 Whole Cell Lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-VWF antigen affinity purified polyclonal antibody (Catalog # PB9062) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for VWF at approximately 309KD. The expected band size for VWF is at 309KD.
VWF was detected in frozen section of human placenta tissue . Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-VWF Antibody (PB9062) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,