Anti-Protein C/PROC Antibody
|Applications||Flow Cytometry, IHC, ICC, WB|
|Product Name||Anti-Protein C/PROC Antibody|
|Storage & Handling||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.|
|Description||Rabbit IgG polyclonal antibody for Vitamin K-dependent protein C(PROC) detection. Tested with WB, IHC-P, IHC-F, ICC, FCM in Human.|
|Cite This Product||Anti-Protein C/PROC Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1682)|
|Contents/Buffer||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Immunogen||A synthetic peptide corresponding to a sequence at the C-terminus of human Protein C(446-461aa HGHIRDKEAPQKSWAP).|
Assay Dilutions Overview
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Western blot, 0.1-0.5μg/ml, Human
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Human
Immunocytochemistry, 0.5-1μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.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), IHC(F) and ICC.
Images And Assay Conditions
Figure 1. Western blot analysis of Protein C using anti- Protein C antibody (PA1682).
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: JURKAT Cell Lysate
Lane 2: CEM Cell Lysate
Lane 3: SMMC Cell Lysate
Lane 4: HELA 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- Protein C antigen affinity purified polyclonal antibody (Catalog # PA1682) at 0.5 Î¼g/mL overnight at 4Â°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 Protein C at approximately 36KD. The expected band size for Protein C is at 52KD.
Figure 3. Flow Cytometry analysis of A549 cells using anti-PROC antibody (PA1682).
Overlay histogram showing A549 cells stained with PA1682 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PROC Antibody (PA1682,1Î¼g/1x106 cells) for 30 min at 20Â°C. DyLightÂ®488 conjugated goat anti-rabbit IgG (BA1127, 5-10Î¼g/1x106 cells) was used as secondary antibody for 30 minutes at 20Â°C. Isotype control antibody (Green line) was rabbit IgG (1Î¼g/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 2. IHC analysis of Protein C using anti- Protein C antibody (PA1682).
Protein C was detected in paraffin-embedded section of human mammary cancer tissues. 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 1Î¼g/ml rabbit anti- Protein C Antibody (PA1682) overnight at 4Â°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37Â°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Vitamin K-dependent protein C|
|Tissue Specificity||Plasma; synthesized in the liver.|
|Alternative Names||Vitamin K-dependent protein C;18.104.22.168;Anticoagulant protein C;Autoprothrombin IIA;Blood coagulation factor XIV;Vitamin K-dependent protein C light chain;Vitamin K-dependent protein C heavy chain;Activation peptide;PROC;|
|Subcellular Localization||Secreted .|
|Molecular Weight||52071 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Protein C is a vitamin K-dependent serine protease that regulates blood coagulation by inactivating factors Va and VIIIa in the presence of calcium ions and phospholipids. .|
*You can search these to find other products in these research areas.
|Background||Protein C(PROC), also called PC, is a zymogenic(inactive) protein, the activated form of which plays an important role in regulating blood clotting, inflammation, cell death and maintaining the permeability of blood vessel walls in humans and other animals. The PROC gene is mapped on 2q14.3. The PROC gene contains 8 exons and spans about 11 kb by Foster et al. The conversion of protein C to a protease with anticoagulant function by thrombin requires as a cofactor thrombomodulin, an endothelial cell membrane protein. Riewald et al. demonstrated that activated protein C uses the endothelial cell protein C receptor as a coreceptor for cleavage of protease-activated receptor-1 on endothelial cells. Faust et al. demonstrated that the endothelial pathways required for protein C activation are impaired in severe meningococcal sepsis. They stated that improvement in the outcome of children with meningococcal sepsis who were treated with unactivated protein C concentrates had been described in case reports and in 1 uncontrolled series.|
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Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugationA: Yes, please contact us at firstname.lastname@example.org for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WBA: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugationA: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact email@example.com
Q: What should I use for negative control?A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signalA: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?A: You can find the immunogen sequence under "
Q: What is the expected band size? Why is it different than the observed band size?A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>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<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>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.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?A: Check our protocols under the tech support tab.