Product Info Summary
SKU: | PA1681 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-Myosin Phosphatase/PPP1R12A Antibody
View all Myosin Phosphatase Antibodies
SKU/Catalog Number
PA1681
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Myosin Phosphatase/PPP1R12A Antibody catalog # PA1681. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Myosin Phosphatase/PPP1R12A Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1681)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the N-terminus of human PPP1R12A, identical to the related rat and mouse sequences.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PA1681 is reactive to PPP1R12A in Human, Mouse, Rat
Applications
PA1681 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee
Observed Molecular Weight
140 kDa
Calculated molecular weight
115.281kDa
Background of Myosin Phosphatase
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.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Assay dilution & Images
Reconsitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
Validation Images & Assay Conditions
Click image to see more details
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.
Click image to see more details
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.
Click image to see more details
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.
Click image to see more details
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.
Click image to see more details
Figure 5. IF analysis of PPP1R12A using anti-PPP1R12A antibody (PA1681).
PPP1R12A was detected in immunocytochemical section of SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-PPP1R12A Antibody (PA1681) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Protein Target Info & Infographic
Gene/Protein Information For PPP1R12A (Source: Uniprot.org, NCBI)
Gene Name
PPP1R12A
Full Name
Protein phosphatase 1 regulatory subunit 12A
Weight
115.281kDa
Alternative Names
MBSmyosin phosphatase, target subunit 1; MGC133042; Myosin phosphatase target subunit 1; Myosin phosphatase-targeting subunit 1; MYPT1protein phosphatase 1 regulatory subunit 12A; protein phosphatase 1, regulatory (inhibitor) subunit 12A; Protein phosphatase myosin-binding subunit PPP1R12A GUBS, M130, MBS, MYPT1 protein phosphatase 1 regulatory subunit 12A protein phosphatase 1 regulatory subunit 12A|myosin binding subunit|myosin phosphatase, target subunit 1|myosin phosphatase-targeting subunit 1|protein phosphatase 1, regulatory (inhibitor) subunit 12A|protein phosphatase myosin-binding subunit
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on PPP1R12A, check out the PPP1R12A Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for PPP1R12A: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Myosin Phosphatase/PPP1R12A Antibody (PA1681)
Hello CJ!
PA1681 has been cited in 1 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
The role of Rho/Rho-kinase pathway and the neuroprotective effects of fasudil in chronic cerebral ischemia
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Customer Q&As
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6 Customer Q&As for Anti-Myosin Phosphatase/PPP1R12A Antibody
Question
Do you have a BSA free version of anti-Myosin Phosphatase/PPP1R12A antibody PA1681 available?
Verified Customer
Verified customer
Asked: 2020-02-26
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-Myosin Phosphatase/PPP1R12A antibody PA1681 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2020-02-26
Question
Would anti-Myosin Phosphatase/PPP1R12A antibody PA1681 work on canine Flow Cytometry with liver?
Verified Customer
Verified customer
Asked: 2019-07-17
Answer
Our lab technicians have not validated anti-Myosin Phosphatase/PPP1R12A antibody PA1681 on canine. You can run a BLAST between canine and the immunogen sequence of anti-Myosin Phosphatase/PPP1R12A antibody PA1681 to see if they may cross-react. If the sequence homology is close, then you can perform a pilot test. Keep in mind that since we have not validated canine samples, this use of the antibody is not covered by our guarantee. However we have an innovator award program that if you test this antibody and show it works in canine liver in Flow Cytometry, you can get your next antibody for free.
Boster Scientific Support
Answered: 2019-07-17
Question
Is this PA1681 anti-Myosin Phosphatase/PPP1R12A antibody reactive to the isotypes of PPP1R12A?
Verified Customer
Verified customer
Asked: 2019-05-31
Answer
The immunogen of PA1681 anti-Myosin Phosphatase/PPP1R12A antibody is 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. Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2019-05-31
Question
Does PA1681 anti-Myosin Phosphatase/PPP1R12A antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2018-12-05
Answer
As indicated on the product datasheet, PA1681 anti-Myosin Phosphatase/PPP1R12A antibody as been validated on WB. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2018-12-05
Question
I was wanting to use your anti-Myosin Phosphatase/PPP1R12A antibody for WB for rat brain liver on frozen tissues, but I want to know if it has been validated for this particular application. Has this antibody been validated and is this antibody a good choice for rat brain liver identification?
Verified Customer
Verified customer
Asked: 2018-10-25
Answer
You can see on the product datasheet, PA1681 anti-Myosin Phosphatase/PPP1R12A antibody has been validated for Flow Cytometry, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in rat brain liver in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2018-10-25
Question
We appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for brain liver using anti-Myosin Phosphatase/PPP1R12A antibody PA1681. Let me know if you need anything else.
L. Parker
Verified customer
Asked: 2013-06-11
Answer
I appreciate the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2013-06-11