Antibody data
- Antibody Data
- Antigen structure
- References [4]
- Comments [0]
- Validations
- Western blot [2]
- Flow cytometry [1]
- Other assay [2]
Submit
Validation data
Reference
Comment
Report error
- Product number
- 44-968G - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-PKC lambda/iota (Thr557, Thr564) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Storage
- -20°C
Submitted references Oncogenic PKC-ι activates Vimentin during epithelial-mesenchymal transition in melanoma; a study based on PKC-ι and PKC-ζ specific inhibitors.
Mature induced-pluripotent-stem-cell-derived human podocytes reconstitute kidney glomerular-capillary-wall function on a chip.
Two novel atypical PKC inhibitors; ACPD and DNDA effectively mitigate cell proliferation and epithelial to mesenchymal transition of metastatic melanoma while inducing apoptosis.
Regulation of polarized morphogenesis by protein kinase C iota in oncogenic epithelial spheroids.
Ratnayake WS, Apostolatos CA, Apostolatos AH, Schutte RJ, Huynh MA, Ostrov DA, Acevedo-Duncan M
Cell adhesion & migration 2018;12(5):447-463
Cell adhesion & migration 2018;12(5):447-463
Mature induced-pluripotent-stem-cell-derived human podocytes reconstitute kidney glomerular-capillary-wall function on a chip.
Musah S, Mammoto A, Ferrante TC, Jeanty SSF, Hirano-Kobayashi M, Mammoto T, Roberts K, Chung S, Novak R, Ingram M, Fatanat-Didar T, Koshy S, Weaver JC, Church GM, Ingber DE
Nature biomedical engineering 2017;1
Nature biomedical engineering 2017;1
Two novel atypical PKC inhibitors; ACPD and DNDA effectively mitigate cell proliferation and epithelial to mesenchymal transition of metastatic melanoma while inducing apoptosis.
Ratnayake WS, Apostolatos AH, Ostrov DA, Acevedo-Duncan M
International journal of oncology 2017 Nov;51(5):1370-1382
International journal of oncology 2017 Nov;51(5):1370-1382
Regulation of polarized morphogenesis by protein kinase C iota in oncogenic epithelial spheroids.
Linch M, Sanz-Garcia M, Rosse C, Riou P, Peel N, Madsen CD, Sahai E, Downward J, Khwaja A, Dillon C, Roffey J, Cameron AJ, Parker PJ
Carcinogenesis 2014 Feb;35(2):396-406
Carcinogenesis 2014 Feb;35(2):396-406
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Lysates prepared from Jurkat cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-4) or treated with lambda phosphatase (5), blocked with a 5% low-fat milk-TBST buffer for one hour at room temperature, and incubated with PKCiota (pT555) antibody for two hours at room temperature in a 3% low-fat milk-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine-containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F (ab’)2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and bands were detected using the Pierce SuperSignal™ method. The data show that the phosphopeptide corresponding to PKCiota (pT555) blocks the antibody signal. The peptide corresponding to PKCzeta (pT560) blocks the antibody signal and the peptides corresponding to PKC isoforms betaI (pT642) and gamma (pT655) partially block the antibody signal (data not shown), suggesting cross-reactivity of the antibody with these sites. The antibody signal was not blocked by the corresponding peptides of any other PKC isoforms. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Lysates prepared from Jurkat cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-4) or treated with lambda phosphatase (5), blocked with a 5% low-fat milk-TBST buffer for one hour at room temperature, and incubated with PKCiota (pT555) antibody for two hours at room temperature in a 3% low-fat milk-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine-containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F (ab’)2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and bands were detected using the Pierce SuperSignal™ method. The data show that the phosphopeptide corresponding to PKCiota (pT555) blocks the antibody signal. The peptide corresponding to PKCzeta (pT560) blocks the antibody signal and the peptides corresponding to PKC isoforms betaI (pT642) and gamma (pT655) partially block the antibody signal (data not shown), suggesting cross-reactivity of the antibody with these sites. The antibody signal was not blocked by the corresponding peptides of any other PKC isoforms. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of PKC-iota [pT555]/ PKC lambda [pT563] was done on HeLa cells treated with PMA (200nM, 20 minutes). 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 PKC-iota [pT555]/ PKC lambda [pT563] Rabbit Polyclonal Antibody (44968G, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) 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
- Fig. 1. Characterization of oncogenic MDCK cells. ( A ) MDCK cell variants cultured in Matrigel (3D) for 6 days alongside phase images of their growth in 2D. Phalloidin (red)-stained actin and Hoechst (blue)-identified nuclei. Scale bar represents 50 mum. Lysates of MDCK oncogenic variant cell lines in log phase growth were immunoblotted for ( B ) defining proteins and ( C ) phospho-PKCiota/total PKCiota.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 4. Effect of inhibitors (ICA-1S, ICA-1T and zeta-Stat) on aPKC expression, apoptosis, and signaling pathways related to EMT in melanoma cells. Expression of the protein levels of phosphorylated PKC-iota, total PKC-iota, phosphorylated PKC-zeta, total PKC-zeta, Caspase-3, cleaved PARP, total PARP, Bcl-2, beta-catenin, Vimentin, phosphorylated Vimentin, Par6, phosphorylated PTEN, RhoA, E-cadherin, phosphorylated AKT and NF-kappaB p65, IkappaB, phosphorylated IkappaB and phosphorylated IKKalpha/beta for the inhibitor treatments (2.5 uM of ICA-1S, 1 muM of ICA-1T and 5 muM of zeta-Stat) are shown in Fig. 4A . 40-80 ug of protein was loaded in to each well and beta-actin was used as the loading control in each Western blot. Fig. 4B represents the densitometry values for Western blots in Fig. 4A . Experiments ( N = 3) were performed in each trial and representative bands are shown.