詳細說明
概述
產(chǎn)品名稱Anti-PKC alpha+beta1+beta2抗體[EPR16794]
描述
兔單克隆抗體[EPR16794] to PKC alpha+beta1+beta2
特異性This antibody also interacts weakly with PKCgamma
經(jīng)測試應用WB,IHC-P,ICC/IF,IP
種屬反應性
與反應: Mouse, Rat, Human
免疫原
Recombinant fragment within Human PKC alpha+beta1+beta2 aa 450 to the C-terminus. The exact sequence is proprietary.
Database link: P05771Run BLAST with
Run BLAST with
陽性對照
WB: Active human PKC alpha full length protein; Active human PKC beta 1 full length protein; Active human PKC beta 2 full length protein; Active human PKC gamma full length protein; Human fetal brain and fetal kidney lysates; Jurkat, SH-SY5Y and L-929 whole cell lysates; Rat and mouse thymus lysates. IHC-P: Human breast cancer, Mouse liver and Rat testis tissues. ICC/IF: HeLa cells. IP: Mouse brain whole cell lysate.
常規(guī)說明
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb? technology. RabMAb? technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
性能
形式Liquid
存放說明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.
存儲溶液Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA純度Protein A purified
克隆單克隆
克隆編號EPR16794
同種型IgG
Anti-PKC alpha+beta1+beta2 antibody [EPR16794] 圖像
Western blot - Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521)
All lanes : Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521) at 1/2000 dilution
Lane 1 : Active human PKC alpha full length protein
Lane 2 : Active human PKC beta 1 full length protein
Lane 3 : Active human PKC beta 2 full length protein
Lane 4 : Active human PKC gamma full length protein
Lane 5 : Active human PKC delta full length protein
Lane 6 : Active human PKC eta full length protein
Lane 7 : Active human PKC epsilon full length protein
Lane 8 : Active human PKC theta full length protein
Lane 9 : Active human PKC mu full length protein
Lysates/proteins at 0.02 μg per lane.
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size : 77, 78 kDa
Exposure time : 5 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
Active human PKC alpha full length protein (Catalog#ab55672) contains aa1-672 with GST-tag; Active human PKC beta 1 full length protein (Catalog#ab60840) contains aa1-671 with GST-tag; Active human PKC beta 2 full length protein (Catalog#ab60841) contains aa1-673 with GST-tag; Active human PKC gamma full length protein (Catalog#ab60842) contains aa1-677 with GST-tag; Active human PKC delta full length protein (Catalog#ab60844) contains aa1-676 with GST-tag; Active human PKC eta full length protein (Catalog#ab60849) contains aa1-683 with GST-tag; Active human PKC epsilon full length protein (Catalog#ab60847) contains aa1-737 with GST-tag; Active human PKC theta full length protein (Catalog#ab56641) contains aa1-706 with GST-tag; Active human PKC mu full length protein (Catalog#ab60873) contains aa1-912 with GST-tag.
Western blot - Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521)
Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521) at 1/2000 dilution + Active human PKC gamma full length protein at 0.02 μg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size : 77, 78 kDa
Exposure time : 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Active human PKC gamma full length protein (Catalog # ab60842) contains aa1-677 with GST-tag.
Western blot - Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521)
All lanes : Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521) at 1/5000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal kidney lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 4 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
Lysates/proteins at 20 μg per lane.
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size : 77, 78 kDa
Observed band size : 78 kDa (why is the actual band size different from the predicted?)
Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521)
All lanes : Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521) at 1/5000 dilution
Lane 1 : L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate
Lane 2 : Rat thymus lysate
Lane 3 : Mouse thymus lysate
Lysates/proteins at 10 μg per lane.
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size : 77, 78 kDa
Observed band size : 78 kDa (why is the actual band size different from the predicted?)
Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PKC alpha+beta1+beta2 with ab179521 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor? 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear and cytoplasm staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor?594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab179521 at 1/100 dilution followed by ab150120 (AlexaFluor?594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor?488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521)
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling PKC alpha+beta1+beta2 with ab179521 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on Human breast cancer is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521)
Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling PKC alpha+beta1+beta2 with ab179521 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse liver is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521)
Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling PKC alpha+beta1+beta2 with ab179521 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on rat testis is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunoprecipitation - Anti-PKC alpha+beta1+beta2 antibody [EPR16794] (ab179521)
PKC alpha+beta1+beta2 was immunoprecipitated from 1mg of Mouse brain whole cell lysate with ab179521 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab179521 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.
Lane 1: Mouse brain whole cell lysate 10μg (Input).
Lane 2: ab179521 IP in Mouse brain whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab179521 in Mouse brain whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.