Anti-PKC iota (phospho T555 + T563)抗体
兔多克隆抗体to PKC iota (phospho T555 + T563)
Rabbit
This antibody reacts with PKC lambda immunoprecipitates, indicating cross-reactivity for PKC lambda [pT563]. PKC zeta [pT560] (83% homologous) has been shown to cross-react by peptide competition. Peptide competition also suggests that this antibody may partially cross-react with PKC beta 1 [pS642] (58% homologous) and PKC nu [pT655] (42% homologous).
适用于: Flow Cyt, WBmore details
与反应: Mouse, Human
Synthetic peptide corresponding to PKC iota (phospho T555 + T563).
WB: Jurkat cells.
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Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA
浓度
50 µl 浓度为 0.9 mg/ml
Immunogen affinity purified
The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated PKC iota. The final product is generated by affinity chromatography using a PKC iota-derived peptide that is phosphorylated at threonine 555.
多克隆
IgG
Abpromise™承诺保证使用ab5813于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| Flow Cyt | Use at an assay dependent concentration. | |
| WB | 1/1000. Detects a band of approximately 76 kDa. |
Entrez Gene: 5584 Human
Entrez Gene: 18759 Mouse
Omim: 600539 Human
SwissProt: P41743 Human
SwissProt: Q62074 Mouse
Unigene: 478199 Human
Unigene: 291554 Mouse
Atypical protein kinase C-lambda/iota antibody
DXS1179E antibody
KPCI_HUMAN antibody
MGC26534 antibody
nPKC iota antibody
nPKC-iota antibody
nPKCiota antibody
PKC lambda antibody
PKC lambda/iota antibody
PKCI antibody
PKCiota antibody
PKClambda antibody
PRKC iota antibody
PRKC lambda/iota antibody
PRKC-lambda/iota antibody
PRKCI antibody
Protein kinase C iota antibody
Protein kinase C iota type antibody
aPKC lambda/iota antibody
aPKC-lambda/iota antibody
Atypical protein kinase C lambda/iota antibody
Western blot - Anti-PKC iota (phospho T555 + T563) antibody (ab5813)
Peptide Competition and Phosphatase Treatment: 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 ab5813 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 and bands were detected using the Pierce SuperSignalTM method. The data show that the phosphopeptide corresponding to PKC iota [pT555] blocks the antibody signal. The peptide corresponding to PKC zeta [pT560] blocks the antibody signal and the peptides corresponding to PKC isoforms beta 1 [pT642] and gamma [pÔ655] 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.
Peptide Competition and Phosphatase Treatment: 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 ab5813 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 and bands were detected using the Pierce SuperSignalTM method. The data show that the phosphopeptide corresponding to PKC iota [pT555] blocks the antibody signal. The peptide corresponding to PKC zeta [pT560] blocks the antibody signal and the peptides corresponding to PKC isoforms beta 1 [pT642] and gamma [pÔ655] 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.
