Antibody data
- Antibody Data
- Antigen structure
- References [5]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Flow cytometry [1]
- Other assay [5]
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Validation data
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- Product number
- 43-8100 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PP1 alpha Monoclonal Antibody (10C6-3)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse, Rat, Canine
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 10C6-3
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references Deficiency of macrophage PHACTR1 impairs efferocytosis and promotes atherosclerotic plaque necrosis.
Deregulated PP1α phosphatase activity towards MAPK activation is antagonized by a tumor suppressive failsafe mechanism.
MYC dephosphorylation by the PP1/PNUTS phosphatase complex regulates chromatin binding and protein stability.
PLEKHA7 defines an apical junctional complex with cytoskeletal associations and miRNA-mediated growth implications.
lncRNA directs cooperative epigenetic regulation downstream of chemokine signals.
Kasikara C, Schilperoort M, Gerlach B, Xue C, Wang X, Zheng Z, Kuriakose G, Dorweiler B, Zhang H, Fredman G, Saleheen D, Reilly MP, Tabas I
The Journal of clinical investigation 2021 Apr 15;131(8)
The Journal of clinical investigation 2021 Apr 15;131(8)
Deregulated PP1α phosphatase activity towards MAPK activation is antagonized by a tumor suppressive failsafe mechanism.
Chen M, Wan L, Zhang J, Zhang J, Mendez L, Clohessy JG, Berry K, Victor J, Yin Q, Zhu Y, Wei W, Pandolfi PP
Nature communications 2018 Jan 15;9(1):159
Nature communications 2018 Jan 15;9(1):159
MYC dephosphorylation by the PP1/PNUTS phosphatase complex regulates chromatin binding and protein stability.
Dingar D, Tu WB, Resetca D, Lourenco C, Tamachi A, De Melo J, Houlahan KE, Kalkat M, Chan PK, Boutros PC, Raught B, Penn LZ
Nature communications 2018 Aug 29;9(1):3502
Nature communications 2018 Aug 29;9(1):3502
PLEKHA7 defines an apical junctional complex with cytoskeletal associations and miRNA-mediated growth implications.
Kourtidis A, Anastasiadis PZ
Cell cycle (Georgetown, Tex.) 2016;15(4):498-505
Cell cycle (Georgetown, Tex.) 2016;15(4):498-505
lncRNA directs cooperative epigenetic regulation downstream of chemokine signals.
Xing Z, Lin A, Li C, Liang K, Wang S, Liu Y, Park PK, Qin L, Wei Y, Hawke DH, Hung MC, Lin C, Yang L
Cell 2014 Nov 20;159(5):1110-1125
Cell 2014 Nov 20;159(5):1110-1125
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on nuclear enriched extracts of HeLa (Lane 1), MCF7 (Lane 2), A549 (Lane 3), PC-3 (Lane 4), Hep G2 (Lane 5), T98G (Lane 6), HEL 92.1.7 (Lane 7), Raji (Lane 8) and THP-1 (Lane 9). The blots were probed with Anti- PP1 Mouse Monoclonal Antibody (Product # 43-8100, 2µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjµgate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A ~ 38 kDa band corresponding to PP1 was observed across all cell lines. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of PP1 alpha was achieved by transfecting HeLa cells with PP1 alpha specific siRNAs (Silencer® select Product # s10930). Western blot analysis (Fig a) was performed using whole cell extracts from the PP1 alpha knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and un-transfected cells (lane 1). The blots were probed with Anti- PP1 alpha Mouse monoclonal Antibody (Product # 43-8100, 2µg/mL) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to PP1 alpha.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PP1 was performed using 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PP1 (10C6-3) Mouse Monoclonal Antibody (Product # 43-8100) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of PP1 was done on MCF7 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with PP1 Mouse Monoclonal Antibody (Product # 43-8100, red histogram) or with mouse isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (Product # A11059) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control..
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
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- Invitrogen Antibodies (provider)
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- Invitrogen Antibodies (provider)
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- Invitrogen Antibodies (provider)
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- Invitrogen Antibodies (provider)
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