Recombinant protein within mouse PKC delta aa 525-671 / 676.
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IF-Tissue, IHC-P
分子量
Predicted band size: 78 kDa
阳性对照
HeLa cell lysate, Jurkat cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, RH-35, mouse brain tissue, rat brain tissue, human tonsil tissue, human liver tissue, human spleen tissue, human breast carcinoma tissue, mouse testis tissue, mouse liver tissue.
Members of the protein kinase C (PKC) family play a key regulatory role in a variety of cellular functions, including cell growth and differentiation, gene expression, hormone secretion and membrane function. PKCs were originally identified as serine/threonine protein kinases whose activity was dependent on calcium and phospholipids. Diacylglycerols (DAG) and tumor promoting phorbol esters bind to and activate PKC. PKCs can be subdivided into at least two major classes, including conventional (c) PKC isoforms (α, βI, βII and γ) and novel (n) PKC isoforms (δ, ε, ζ, η, θ, λ/ι, μ and ν). Patterns of expression for each PKC isoform differ among tissues and PKC family members exhibit clear differences in their cofactor dependencies. For instance, the kinase activities of PKC δ and ε are independent of Ca2+. On the other hand, most of the other PKC members possess phorbol ester-binding activities and kinase activities.
背景文献
1. Slusarczyk J et al. Anti-inflammatory properties of tianeptine on lipopolysaccharide-induced changes in microglial cells involve toll-like receptor-related pathways. J Neurochem 136:958-70 (2016).
2. Song X et al. RasGRPs are targets of the anti-cancer agent ingenol-3-angelate. PLoS One 8:e72331 (2013).
序列相似性
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily.
翻译后修饰
Autophosphorylated and/or phosphorylated at Thr-507, within the activation loop; phosphorylation at Thr-507 is not a prerequisite for enzymatic activity. Autophosphorylated at Ser-299, Ser-302 and Ser-304. Upon TNFSF10/TRAIL treatment, phosphorylated at Tyr-155; phosphorylation is required for its translocation to the endoplasmic reticulum and cleavage by caspase-3. Phosphorylated at Tyr-313, Tyr-334 and Tyr-567; phosphorylation of Tyr-313 and Tyr-567 following thrombin stimulation potentiates its kinase activity. Phosphorylated by protein kinase PDPK1; phosphorylation is inhibited by the apoptotic C-terminal cleavage product of PKN2.; Proteolytically cleaved into a catalytic subunit and a regulatory subunit by caspase-3 during apoptosis which results in kinase activation.
Western blot analysis of PKC delta on different lysates with Rabbit anti-PKC delta antibody (ET1701-85) at 1/5,000 dilution.
Lane 1: HeLa cell lysate (15 µg/Lane) Lane 2: Jurkat cell lysate (15 µg/Lane) Lane 3: Mouse brain tissue lysate (20 µg/Lane) Lane 4: Rat brain tissue lysate (20 µg/Lane)
Predicted band size: 78 kDa Observed band size: 78 kDa
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-85) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of RH-35 cells labeling PKC delta with Rabbit anti-PKC delta antibody (ET1701-85) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-PKC delta antibody (ET1701-85) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-PKC delta antibody (ET1701-85) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-85) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-PKC delta antibody (ET1701-85) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-85) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-PKC delta antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-85, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-PKC delta antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-85, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-PKC delta antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-85, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-PKC delta antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-85, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-PKC delta antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-85, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-PKC delta antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-85, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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