重组Anti-PKC delta抗体[EPR17075] (ab182126)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17075] to PKC delta
- Suitable for: Flow Cyt, IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
概述
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产品名称
Anti-PKC delta抗体[EPR17075]
参阅全部 PKC delta 一抗 -
描述
兔单克隆抗体[EPR17075] to PKC delta -
宿主
Rabbit -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P MouseRatHumanWB MouseRatHuman -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HEK-239T, A431, C6, NIH/3T3, HAP1 and HeLa whole cell lysates, human fetal brain and fetal heart, mouse and rat thymus and brain and rat spleen tissue lysates. IHC-P: Human spleen, human transitional cell carcinoma of bladder, mouse liver and rat testis tissues. ICC/IF: Wild-type HAP1 (PMA-treated and untreated) and HeLa cells. Flow Cyt: HeLa cells.
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常规说明
Protein kinase C delta type (PKCD) is an enzyme encoded for by the PRKCD gene and is expressed ubiquitously among cells and tissues. It is thought that PKCD, when activated in distinct ways, plays critical roles in cellular functions such as the control of growth, differentiation and apoptosis.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR17075 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
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Recombinant Protein
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab182126 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
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Flow Cyt |
Human
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ICC/IF |
Human
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IHC-P |
Mouse
Rat
Human
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WB |
Mouse
Rat
Human
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应用 | Ab评论 | 说明 |
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Flow Cyt |
1/250.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-P | (4) |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB | (1) |
1/5000. Detects a band of approximately 40, 78 kDa (predicted molecular weight: 78 kDa).
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ICC/IF |
Use a concentration of 5 µg/ml.
Treatment with 10nM PMA for 10 min induces translocation of PKCδ to the membrane. |
说明 |
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Flow Cyt
1/250. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/5000. Detects a band of approximately 40, 78 kDa (predicted molecular weight: 78 kDa). |
ICC/IF
Use a concentration of 5 µg/ml. Treatment with 10nM PMA for 10 min induces translocation of PKCδ to the membrane. |
靶标
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功能
This is calcium-independent, phospholipid-dependent, serine- and threonine-specific enzyme. PKC is activated by diacylglycerol which in turn phosphorylates a range of cellular proteins. PKC also serves as the receptor for phorbol esters, a class of tumor promoters. May play a role in antigen-dependent control of B-cell function. Phosphorylates MUC1 in the C-terminal and regulates the interaction between MUC1 and beta-catenin. -
序列相似性
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 C2 domain.
Contains 2 phorbol-ester/DAG-type zinc fingers.
Contains 1 protein kinase domain. -
结构域
The C1 domain, containing the phorbol ester/DAG-type region 1 (C1A) and 2 (C1B), is the diacylglycerol sensor.
The C2 domain is a non-calcium binding domain. It binds proteins containing phosphotyrosine in a sequence-specific manner. -
翻译后修饰
Phosphorylated on Thr-507, within the activation loop. Autophosphorylated and/or phosphorylated. Although the Thr-507 phosphorylation occurs it is not a prerequisite for enzymatic activity. -
细胞定位
Cytoplasm. Membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 5580 Human
- Entrez Gene: 18753 Mouse
- Entrez Gene: 170538 Rat
- Omim: 176977 Human
- SwissProt: Q05655 Human
- SwissProt: P28867 Mouse
- SwissProt: P09215 Rat
- Unigene: 155342 Human
see all -
别名
- CVID9 antibody
- D14Ertd420e antibody
- Kinase PKC delta antibody
see all
图片
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All lanes : Anti-PKC delta antibody [EPR17075] (ab182126) at 1/5000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PRKCD knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 78 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab182126 observed at 80 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab182126 was shown to react with PKC delta in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265721 (knockout cell lysate ab257043) was used. Wild-type HeLa and PKC delta knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab182126 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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ab182126 staining PKCδ in untreated wild-type HAP1 cells (top panel) and PKCδ untreated knockout HAP1 cells (bottom panel). Untreated cells show PKCδ being expressed in the cytoplasm. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab182126 at 1/200 dilution and ab7291 at 1ug/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150117) at 2ug/ml (shown in pseudo-color red). Nuclear DNA was labelled in blue with DAPI.
This product also gave a positive signal under the same testing conditions in HAP1 cells fixed with 100% methanol (5 min).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC delta antibody [EPR17075] (ab182126)
Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling PKC delta with ab182126 at 1/2000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic and weak nuclear staining on cells in the seminiferous tubule of rat testis is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-PKC delta antibody [EPR17075] (ab182126) at 1/5000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : PKC delta knockout HAP1 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 78 kDaLanes 1 - 4: Merged signal (red and green). Green - ab182126 observed at 78 kDa. Red - loading control, ab8226, observed at 42 kDa.
ab182126 was shown to specifically react with PKC delta when PKC delta knockout samples were used. Wild-type and PKC delta knockout samples were subjected to SDS-PAGE. ab182126 and ab8226 (loading control to beta Actin) were diluted to 1/5000 and 1/1000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
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All lanes : Anti-PKC delta antibody [EPR17075] (ab182126) at 1/5000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : PRKCD Knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 78 kDa
Observed band size: 78 kDaLanes 1 - 2: Merged signal (red and green). Green - ab182126 observed at 78 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab182126 was shown to react with PKC in wild-type HEK-293T cells in western blot. The bands observed in PRKCD knockout cell line ab266143 (PRKCD knockout cell lysate ab257042) below 78kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and PRKCD knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab182126 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4
176;
® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging. -
Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling PKC delta with purified ab182126 at 1/250 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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ab182126 staining PKCδ in 10nM PMA-treated wild-type HAP1 cells (top panel) and PKCδ in 10nM PMA-treated knockout HAP1 cells (bottom panel). The cells were treated with 10nM PMA for 10 minutes to induce translocation of PKCδ to the cell membrane. The cells were then fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab182126 at 1/200 dilution and ab7291 at 1ug/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150117) at 2ug/ml (shown in pseudo-color red). Nuclear DNA was labelled in blue with DAPI.
This product also gave a positive signal under the same testing conditions in HAP1 cells fixed with 100% methanol (5 min).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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All lanes : Anti-PKC delta antibody [EPR17075] (ab182126) at 1/5000 dilution
Lane 1 : Mouse brain lysates
Lane 2 : Rat brain lysates
Lane 3 : Rat spleen lysates
Lane 4 : C6 (Rat glial tumor cells) whole cell lysates
Lane 5 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 78 kDaThe 40kDa band represents the cleaved kinase domain.
Blocking/dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC delta antibody [EPR17075] (ab182126)
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling PKC delta with ab182126 at 1/2000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining on kupffer cells of Mouse liver is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-PKC delta antibody [EPR17075] (ab182126) at 1/5000 dilution
Lane 1 : Human fetal brain lysates
Lane 2 : Human fetal heart lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 78 kDaThe 40kDa band represents the cleaved kinase domain.
Blocking/dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC delta antibody [EPR17075] (ab182126)
Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma of bladder tissue labeling PKC delta with ab182126 at 1/2000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic and weak nuclear staining on cancer cells of bladder transitional cell carcinoma is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PKC delta with ab182126 at 1/100 dilution. The cells were permeabilised with 0.1% Triton X-100. Goat anti-rabbit IAlexa Fluor® 488 (IgG) (ab150077) at 1/400 dilution was used as the secondary antibody (green). The confocal image shows both cytoplasmic and nuclear staining on HeLa cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and ab150120 (goat anti-mouse AlexaFluor® 594 secondary antibody) at 1/500 dilution (red).
The negative controls are as follows;
1. ab182126 at 1/100 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution. -
All lanes : Anti-PKC delta antibody [EPR17075] (ab182126) at 1/10000 dilution
Lane 1 : A431 (Human epidermoid carcinoma) whole cell lysates at 10 µg
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates at 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 78 kDaThe 40kDa band represents the cleaved kinase domain.
Blocking/dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC delta antibody [EPR17075] (ab182126)
Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling PKC delta with ab182126 at 1/2000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic and nucleus staining on lymphocytes of Human spleen is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-PKC delta antibody [EPR17075] (ab182126) at 1/100000 dilution
Lane 1 : Mouse thymus lysates
Lane 2 : Rat thymus lysates
Lysates/proteins at 10 mg/ml per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 78 kDaThe 40kDa band represents the cleaved kinase domain.
Blocking/dilution buffer: 5% NFDM/TBST.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (24)
ab182126 被引用在 24 文献中.
- Du J et al. MicroRNA-26a/b have protective roles in oral lichen planus. Cell Death Dis 11:15 (2020). PubMed: 31907356
- Ma JX et al. Couplet medicines of leech and centipede granules improve erectile dysfunction via inactivation of the CaSR/PLC/PKC signaling in streptozotocin-induced diabetic rats. Biosci Rep 40:N/A (2020). PubMed: 31922200
- Lenkavska L et al. Searching for combination therapy by clustering methods: Stimulation of PKC in Golgi apparatus combined with hypericin induced PDT. Photodiagnosis Photodyn Ther N/A:101813 (2020). PubMed: 32442674
- Li S et al. PKC-d deficiency in B cells displays osteopenia accompanied with upregulation of RANKL expression and osteoclast-osteoblast uncoupling. Cell Death Dis 11:762 (2020). PubMed: 32938907
- Lee S et al. Docosahexaenoic Acid Inhibits Expression of Fibrotic Mediators in Mice With Chronic Pancreatitis. J Cancer Prev 24:233-239 (2019). PubMed: 31950023