Anti-Myosin Phosphatase/PPP1R12A Antibody

SKU PA1681
Size 100μg/vial
Reactivity Human, Mouse, Rat
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications Flow Cytometry, WB

Overview

Product Name Anti-Myosin Phosphatase/PPP1R12A Antibody
SKU/Catalog Number PA1681
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.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for Protein phosphatase 1 regulatory subunit 12A(PPP1R12A) detection. Tested with WB, FCM in Human;Mouse;Rat.
Cite This Product Anti-Myosin Phosphatase/PPP1R12A Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1681)
Host Rabbit
Contents/Buffer Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
Form Lyophilized
Immunogen A synthetic peptide corresponding to a sequence at the N-terminus of human PPP1R12A (1-17aa MKMADAKQKRNEQLKRW), identical to the related rat and mouse sequences.
Reactivity Human, Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
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.

Images And Assay Conditions

Figure 2. Flow Cytometry analysis of Hela cells using anti-PPP1R12A antibody (PA1681 ).
Overlay histogram showing Hela cells stained with PA1681 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (PA1681 ,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 3. Flow Cytometry analysis of U251 cells using anti-PPP1R12A antibody (PA1681 ).
Overlay histogram showing U251 cells stained with PA1681 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (PA1681 ,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 1. Western blot analysis of PPP1R12A using anti- PPP1R12A antibody (PA1681).
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: human Hela whole cell lysates,
Lane 2: human Jurka7 whole cell lysates,
Lane 3: human HEK293 whole cell lysates,
Lane 4: monkey COS-7 whole cell lysates,
Lane 5: human Raji whole cell lysates,
Lane 6: human K562 whole cell lysates,
Lane 7: human Caco-2 whole cell lysates,
Lane 8: human HepG2 whole cell lysates,
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- PPP1R12A antigen affinity purified polyclonal antibody (Catalog # PA1681) 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 PPP1R12A at approximately 140KD. The expected band size for PPP1R12A is at 115KD.

Figure 4. Western blot analysis of PPP1R12A using anti- PPP1R12A antibody (PA1681).
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: rat brain tissue lysates,
Lane 2: rat C6 whole cell lysates,
Lane 3: mouse liver tissue lysates,
Lane 4: mouse NIH/3T3 whole cell lysates,
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- PPP1R12A antigen affinity purified polyclonal antibody (Catalog # PA1681) 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 PPP1R12A at approximately 140KD. The expected band size for PPP1R12A is at 115KD.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id O14974
Gene Name PPP1R12A
Protein Name Protein phosphatase 1 regulatory subunit 12A
Tissue Specificity Expressed in striated muscles, specifically in type 2a fibers (at protein level). .
Alternative Names Protein phosphatase 1 regulatory subunit 12A;Myosin phosphatase-targeting subunit 1;Myosin phosphatase target subunit 1;Protein phosphatase myosin-binding subunit;PPP1R12A;MBS, MYPT1;
Subcellular Localization Cytoplasm . Along actomyosin filaments and stress fibers.
Molecular Weight 115281 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function Key regulator of protein phosphatase 1C (PPP1C). Mediates binding to myosin. As part of the PPP1C complex, involved in dephosphorylation of PLK1. Capable of inhibiting HIF1AN- dependent suppression of HIF1A activity. .
Research Areas Human, Mouse, Rat

*You can search these to find other products in these research areas.
Background PPP1R12A(Protein phosphatase 1 regulatory subunit 12A), also called MYPT1(Myosin phosphatase target subunit 1), is an enzyme that in humans is encoded by the PPP1R12A gene. PPP1R12A is one of the subunits of myosin phosphatase. Sequencing analysis showed that human PPP1R12A contains 1,030 amino acids with a calculated molecular mass of approximately 115 kD. The PPP1R12A gene is mapped on 12q21.2-q21.3. PPP1R12A is the protein that regulates PP1 function in smooth muscle relaxation. The cellular MYPT1-PP1-delta -specific inhibitor CPI17 caused a loss of merlin function characterized by merlin phosphorylation, Ras activation, and transformation. Jin et al. concluded that PPP1R12A and its substrate merlin are part of a previously undescribed tumor suppressor cascade that can be hindered in 2 ways, by mutation of the NF2 gene and by upregulation of the oncoprotein CPI17.

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Rabbit IgG polyclonal antibody for Protein phosphatase 1 regulatory subunit 12A(PPP1R12A) detection. Tested with WB, FCM in Human;Mouse;Rat.
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PA1681
Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)
$240.00

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Customer Q&As

  • Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
    A: Yes, please contact us at support@bosterbio.com 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, WB
    A: 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: conjugation
    A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.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 signal
    A: 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.
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