Western Blot (WB)
(Figure 1. Western blot analysis of PPP1R12A using anti-PPP1R12A antibody (MBS177838).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: Mouse Skeletal Muscle Tissue Lysate,Lane 2: HELA 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-PPP1R12A antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for PPP1R12A at approximately 150KD. The expected band size for PPP1R12A is at 115KD. )
anti-Human, Mouse PPP1R12A Polyclonal Antibody | anti-PPP1R12A antibody
Anti-PPP1R12A Antibody
Immunohistochemistry (IHC) Paraffin Concentration: 0.5-1ug/ml
Western Blot (WB)
(Figure 1. Western blot analysis of PPP1R12A using anti-PPP1R12A antibody (MBS177838).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: Mouse Skeletal Muscle Tissue Lysate,Lane 2: HELA 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-PPP1R12A antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for PPP1R12A at approximately 150KD. The expected band size for PPP1R12A is at 115KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of PPP1R12A using anti-PPP1R12A antibody (MBS177838).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: Mouse Skeletal Muscle Tissue Lysate,Lane 2: HELA 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-PPP1R12A antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for PPP1R12A at approximately 150KD. The expected band size for PPP1R12A is at 115KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of PPP1R12A using anti-PPP1R12A antibody (MBS177838).PPP1R12A was detected in paraffin-embedded section of Human Glioma 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 1ug/ml rabbit anti-PPP1R12A Antibody (MBS177838) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of PPP1R12A using anti-PPP1R12A antibody (MBS177838).PPP1R12A was detected in paraffin-embedded section of Human Glioma 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 1ug/ml rabbit anti-PPP1R12A Antibody (MBS177838) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of Hela cells using anti-PPP1R12A antibody (MBS177838).Overlay histogram showing Hela cells stained with MBS177838 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (MBS177838,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of Hela cells using anti-PPP1R12A antibody (MBS177838).Overlay histogram showing Hela cells stained with MBS177838 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (MBS177838,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 4. Flow Cytometry analysis of SiHa cells using anti-PPP1R12A antibody (MBS177838).Overlay histogram showing SiHa cells stained with MBS177838 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (MBS177838,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 4. Flow Cytometry analysis of SiHa cells using anti-PPP1R12A antibody (MBS177838).Overlay histogram showing SiHa cells stained with MBS177838 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (MBS177838,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 5. Flow Cytometry analysis of U251 cells using anti-PPP1R12A antibody (MBS177838).Overlay histogram showing U251 cells stained with MBS177838 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (MBS177838,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 5. Flow Cytometry analysis of U251 cells using anti-PPP1R12A antibody (MBS177838).Overlay histogram showing U251 cells stained with MBS177838 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (MBS177838,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
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 two ways, by mutation of the NF2 gene and by upregulation of the oncoprotein CPI17.
NCBI and Uniprot Product Information
NCBI Description
Myosin phosphatase target subunit 1, which is also called the myosin-binding subunit of myosin phosphatase, is one of the subunits of myosin phosphatase. Myosin phosphatase regulates the interaction of actin and myosin downstream of the guanosine triphosphatase Rho. The small guanosine triphosphatase Rho is implicated in myosin light chain (MLC) phosphorylation, which results in contraction of smooth muscle and interaction of actin and myosin in nonmuscle cells. The guanosine triphosphate (GTP)-bound, active form of RhoA (GTP.RhoA) specifically interacted with the myosin-binding subunit (MBS) of myosin phosphatase, which regulates the extent of phosphorylation of MLC. Rho-associated kinase (Rho-kinase), which is activated by GTP. RhoA, phosphorylated MBS and consequently inactivated myosin phosphatase. Overexpression of RhoA or activated RhoA in NIH 3T3 cells increased phosphorylation of MBS and MLC. Thus, Rho appears to inhibit myosin phosphatase through the action of Rho-kinase. Several transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jan 2009]
Uniprot Description
MYPT1: myosin phosphatase target subunit 1, a regulatory subunit of protein phosphatase 1. Also called the myosin-binding subunit of myosin phosphatase. Myosin phosphatase regulates the interaction of actin and myosin downstream of the guanosine triphosphatase Rho. Four splice-variant isoforms have been described.
Protein type: Motility/polarity/chemotaxis; Protein phosphatase, regulatory subunit
Chromosomal Location of Human Ortholog: 12q15-q21
Cellular Component: A band; actin cytoskeleton; centrosome; contractile fiber; cytoplasm; cytosol; focal adhesion; kinetochore; nucleoplasm; Z disc
Molecular Function: enzyme inhibitor activity; phosphatase regulator activity; phosphoprotein phosphatase activity; protein binding; protein kinase binding; signal transducer activity
Biological Process: centrosome organization and biogenesis; G2/M transition of mitotic cell cycle; mitosis; negative regulation of catalytic activity; positive regulation of transcription from RNA polymerase II promoter; protein amino acid dephosphorylation; regulation of cell adhesion; regulation of nucleocytoplasmic transport; signal transduction
Research Articles on PPP1R12A
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Product Notes
The PPP1R12A ppp1r12a (Catalog #AAA177838) is an Antibody and is intended for research purposes only. The product is available for immediate purchase. The Anti-PPP1R12A Antibody reacts with Human, Mouse and may cross-react with other species as described in the data sheet. AAA Biotech's PPP1R12A can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC) Paraffin. Western Blot Concentration: 0.1-0.5ug/ml Immunohistochemistry (IHC) Paraffin Concentration: 0.5-1ug/ml. Researchers should empirically determine the suitability of the PPP1R12A ppp1r12a for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "PPP1R12A, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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