Product Info Summary
SKU: | PB9737 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Monkey, Mouse, Rat |
Host: | Rabbit |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Myosin Phosphatase/PPP1R12A Antibody Picoband™
View all Myosin Phosphatase Antibodies
SKU/Catalog Number
PB9737
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Myosin Phosphatase/PPP1R12A Antibody Picoband™ catalog # PB9737. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Monkey, 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 Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9737)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 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 mouse and rat 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
PB9737 is reactive to PPP1R12A in Human, Monkey, Mouse, Rat
Applications
PB9737 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
130 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 two 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.
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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, Monkey
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
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 (PB9737).
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 Jurkat 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 # PB9737) 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 1:5000 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 130KD. The expected band size for PPP1R12A is at 130KD.
Click image to see more details
Figure 2. IHC analysis of PPP1R12A using anti-PPP1R12A antibody (PB9737).
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 1μg/ml rabbit anti-PPP1R12A Antibody (PB9737) 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.
Click image to see more details
Figure 3. Flow Cytometry analysis of Hela cells using anti-PPP1R12A antibody (PB9737).
Overlay histogram showing Hela cells stained with PB9737 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (PB9737,1μg/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. Flow Cytometry analysis of SiHa cells using anti-PPP1R12A antibody (PB9737).
Overlay histogram showing SiHa cells stained with PB9737 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (PB9737,1μg/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 5. Flow Cytometry analysis of U251 cells using anti-PPP1R12A antibody (PB9737).
Overlay histogram showing U251 cells stained with PB9737 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (PB9737,1μg/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 6. IHC analysis of PPP1R12A using anti-PPP1R12A antibody (PB9737).
PPP1R12A was detected in immunocytochemical section of SiHa cell. 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 (PB9737) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using DyLight®488 Conjugated Avidin (BA1128). Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 7. IHC analysis of PPP1R12A using anti-PPP1R12A antibody (PB9737).
PPP1R12A was detected in immunocytochemical section of U251 cell. 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 (PB9737) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using DyLight®488 Conjugated Avidin (BA1128). Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 8. IHC analysis of PPP1R12A using anti-PPP1R12A antibody (PB9737).
PPP1R12A was detected in immunocytochemical section of U251 cell. 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 (PB9737) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using DyLight®488 Conjugated Avidin (BA1128). Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 9. IHC analysis of PPP1R12A using anti-PPP1R12A antibody (PB9737).
PPP1R12A was detected in immunocytochemical section of U251 cell. 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 (PB9737) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using DyLight®488 Conjugated Avidin (BA1128). Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 10. IHC analysis of PPP1R12A using anti-PPP1R12A antibody (PB9737).
PPP1R12A was detected in immunocytochemical section of U251 cell. 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 (PB9737) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using DyLight®488 Conjugated Avidin (BA1128). Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 11. Western blot analysis of PPP1R12A using anti-PPP1R12A antibody (PB9737).
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 lung tissue lysates,
Lane 3: rat liver tissue lysates,
Lane 4: rat C6 whole cell lysates,
Lane 5: mouse brain tissue lysates,
Lane 6: mouse lung tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: 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 # PB9737) 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 1:5000 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 130KD. The expected band size for PPP1R12A is at 130KD.
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 Picoband™ (PB9737)
Hello CJ!
PB9737 has been cited in 2 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
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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1 Customer Q&As for Anti-Myosin Phosphatase/PPP1R12A Antibody Picoband™
Question
We are currently using anti-Myosin Phosphatase/PPP1R12A antibody PB9737 for mouse tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on goat tissues as well?
Verified Customer
Verified customer
Asked: 2019-08-16
Answer
The anti-Myosin Phosphatase/PPP1R12A antibody (PB9737) has not been tested for cross reactivity specifically with goat tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in goat you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-08-16