详细说明
概述
产品名称Anti-DUSP6抗体[EPR129Y]
描述
兔单克隆抗体[EPR129Y] to DUSP6
经测试应用IHC-P,IHC-FoFr,WB,IP,Flow Cyt,ICC/IF
种属反应性
与反应: Mouse, Rat, Human
免疫原
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human DUSP6 aa 350 to the C-terminus (C terminal).
Database link: Q16828
(Peptide available as ab171765)阳性对照
WB: 3T3 cell lysate IHC: Human gastric carcinoma tissue.
常规说明
This product is a recombinant rabbit monoclonal antibody.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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.
Alternative versions available:
Anti-DUSP6 antibody (Alexa Fluor® 488) [EPR129Y] (ab200750)
Anti-DUSP6 antibody (Alexa Fluor® 647) [EPR129Y] (ab200751)
性能
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. Avoid freeze / thaw cycle.
存储溶液pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA纯度Protein A purified
克隆单克隆
克隆编号EPR129Y
同种型IgG
研究领域
Neuroscience
Neurology process
Neural Signal Transduction
Anti-DUSP6 antibody [EPR129Y] 图像
Western blot - Anti-DUSP6 antibody [EPR129Y] (ab76310)
Anti-DUSP6 antibody [EPR129Y] (ab76310) at 1/5000 dilution (purified) + HepG2 cell lysate at 20 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size : 42 kDa
Observed band size : 42,44 kDa (why is the actual band size different from the predicted?)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBSTab76310 detects an unspecific band around 100 kDa in human materials.
Western blot - Anti-DUSP6 antibody [EPR129Y] (ab76310)
All lanes : Anti-DUSP6 antibody [EPR129Y] (ab76310) at 1/1000 dilution (purified)
Lane 1 : rat brain lysate
Lane 2 : NIH/3T3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size : 42 kDa
Observed band size : 42,44 kDa (why is the actual band size different from the predicted?)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBSTWestern blot - Anti-DUSP6 antibody [EPR129Y] (ab76310)
Anti-DUSP6 antibody [EPR129Y] (ab76310) at 1/1000 dilution (purified) + mouse brain at 10 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size : 42 kDa
Observed band size : 42,44 kDa (why is the actual band size different from the predicted?)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBSTWestern blot - DUSP6 antibody [EPR129Y] (ab76310)
Anti-DUSP6 antibody [EPR129Y] (ab76310) at 1/500 dilution (unpurified) + 3T3 cell lysate at 10 µg
Secondary
HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size : 42 kDa
Observed band size : 42/44 kDa (why is the actual band size different from the predicted?)
Western blot - Anti-DUSP6 antibody [EPR129Y] (ab76310) This image was taken from an abreview by Graeme Stewart.
All lanes : Anti-DUSP6 antibody [EPR129Y] (ab76310) at 1/4000 dilution (unpurified)
Lane 1 : Marker
Lane 2 : Lysate from wild type primary murine embryonic fibroblasts (MEFs) untreated at 10 µg
Lane 3 : Lysate from wild type primary murine embryonic fibroblasts (MEFs) treated with 100ngµl-1, 12-O-tetradecanoylphorbol-13-acetate (TPA) for 2 hours at 10 µg
Lane 4 : Lysate from MKP-3 null primary murine embryonic fibroblasts (MEFs) untreated at 10 µg
Lane 5 : Lysate from MKP-3 null primary murine embryonic fibroblasts (MEFs) treated with 100ngµl-1, 12-O-tetradecanoylphorbol-13-acetate (TPA) for 2 hours at 10 µg
Secondary
Goat anti Rabbit HRP conjugate at 1/10000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 42 kDa
Observed band size : 42 kDa
Exposure time : 2 minutesThis image was taken from an abreview by Graeme Stewart.
See Abreview
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DUSP6 antibody [EPR129Y] (ab76310)
Immunohistochemical staining of paraffin embedded human pancreas with purified ab76310 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - DUSP6 antibody [EPR129Y] (ab76310)
Immunohistochemical staining of paraffin-embedded human gastric carcinoma using unpurified ab76310 at 1/50 dilution.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-DUSP6 antibody [EPR129Y] (ab76310) This image is courtesy of an Abreview submitted by Carlos Sindreu
Immunohistochemistry (PFA perfusion fixed frozen sections) analysis of Mouse brain tissue sections labelling DUSP6 with unpurified ab76310 at 1/400 dilution for 14 hours at 4°C. A biotinylated polyclonal anti-rabbit IgG was used as the secondary antibody at 1/250 dilution.
See Abreview
Immunocytochemistry/ Immunofluorescence - Anti-DUSP6 antibody [EPR129Y] (ab76310)
Immunofluorescence staining of PC-12 cells with purified ab76310 at a working dilution of 1/200, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab76310 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Immunocytochemistry/ Immunofluorescence - DUSP6 antibody [EPR129Y] (ab76310)
ICC/IF image of unpurified ab76310 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76310, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunoprecipitation - Anti-DUSP6 antibody [EPR129Y] (ab76310)
ab76310 (purified) at 1/20 immunoprecipitating DUSP6 in 10 μg NIH-3T3 (Lanes 1 and 2, observed at 42 and 44 kDa). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
Flow Cytometry - Anti-DUSP6 antibody [EPR129Y] (ab76310)
Overlay histogram showing NIH-3T3 cells fixed in 4% PFA and stained with purified ab76310 at a dilution of 1 in 200 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
Flow Cytometry-DUSP6 antibody [EPR129Y](ab76310)
Overlay histogram showing HeLa cells stained with unpurified ab76310 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76310, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized with 0.1% PBS-Tween 20 used under the same conditions.