Anti-JNK1 + JNK2 (phospho T183 + Y185)抗体
参阅全部 JNK1 + JNK2 一抗
兔多克隆抗体to JNK1 + JNK2 (phospho T183 + Y185)
Rabbit
Phosphorylation site-specific antibody selective for the dually phosphorylated form of the c-Jun N-terminal Kinase (JNK)/Stress-Activated Protein Kinase (SAPK) enzymes containing a phosphate on threonine 183 and tyrosine 185 (human JNK 1 + 2). The antibody has been shown to recognize the endogenous, active forms of JNK 1 + 2 in a variety of cell types following treatment by a broad range of extracellular stimuli [e.g. including 293 cells (human embryonic kidney; +/- ultraviolet light) and PC12 cells (rat pheochromocytoma; +/- sorbital)]. The region of JNK1 and JNK2 surrounding T183 + Y185 has a high degree of similarity to the corresponding regions in JNK3 and thus may cross react with this protein if phosphorylated on the corresponding residues.
适用于: ICC/IF, WBmore details
与反应: Mouse, Human
预测可用于: a wide range of other species
Synthetic peptide corresponding to Human JNK1 + JNK2 (phospho T183 + Y185).
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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, 50% Glycerol, 0.1% BSA
BSA is IgG and protease free
浓度
批次浓度范围 50 µl 浓度为 0.75 - 0.9 mg/ml
Immunogen affinity purified
Purified from rabbit serum by sequential epitope specific chromatography. 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 JNK enzymes. The final product is generated by affinity chromatography using a JNK-derived peptide that is phosphorylated at threonine 183 and tyrosine 185, within the activation loop. Note: It is the dually phosphorylated form of these enzymes that has full enzymatic activity.
多克隆
IgG
Abpromise™承诺保证使用ab4821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 |
|---|---|---|
| ICC/IF | (1) | 1/250. 1/100. |
| WB | (7) | 1/1000. Predicted molecular weight: 49, 55 kDa. Band at ~49 kDa represents Jnk1, while the band at ~55 kDa represents Jnk2 |
Entrez Gene: 5599 Human
Entrez Gene: 5601 Human
Entrez Gene: 26419 Mouse
Entrez Gene: 26420 Mouse
Omim: 601158 Human
Omim: 602896 Human
SwissProt: P45983 Human
SwissProt: P45984 Human
SwissProt: Q91Y86 Mouse
SwissProt: Q9WTU6 Mouse
Unigene: 138211 Human
Unigene: 21495 Mouse
c jun N terminal kinase 2 antibody
c-Jun N-terminal kinase 1 antibody
cJun N terminal kinase 1 antibody
JNK 1 antibody
JNK 2 antibody
JNK-46 antibody
JNK2ALPHA antibody
JNK2BETA antibody
MAP kinase 8 antibody
MAP kinase 9 antibody
MAPK 8 antibody
mapk8 antibody
MAPK9 antibody
Mitogen activated protein kinase 8 antibody
Mitogen activated protein kinase 9 antibody
Mitogen-activated protein kinase 8 antibody
MK08_HUMAN antibody
PRKM 8 antibody
PRKM 9 antibody
Stress-activated protein kinase 1 antibody
Stress-activated protein kinase JNK1 antibody
Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
All lanes : Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821) at 1/1000 dilution
Lane 1 : HEK-293 cell line
Lane 2 : HEK-293 treated for 5 minutes with 200 mM of Anisomycin
Lane 3 : HEK-293 treated for 20 minutes with UV
Lane 4 : MCF7 cell line
Lane 5 : MCF7 treated for 5 minutes with 200 mM of Anisomycin
Lane 6 : K562 cell line
Lane 7 : K562 treated for 20 minutes with UV
Lane 8 : HeLa cell line
Lane 9 : HeLa treated for 20 minutes with UV
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/5000 dilution
Predicted band size: 49, 55 kDa
Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature.
Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
MEF1 cells were incubated at 37°C for 48h with vehicle control (0 µM) and 5 µM of glibenclamide (ab120267) in DMSO. Increased expression of of JNK1+JNK2 (phospho T183 + Y185) (ab4821) correlates with an increase in glibenclamide concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated with ab4821 at 1/1000 dilution and ab85139 at 1 µg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.
Immunocytochemistry/ Immunofluorescence - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
ab4821 staining JNK1 + JNK2 (phospho T183 + Y185) in A549 cells (green, panel a) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde, permeabilized with 0.25% Triton X-100 and blocked with 5% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (2ug/ml in 1% BSA) for 3 hours at room temperature. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody (1/400). Nuclei stained with DAPI (blue, panel b), F-actin stained with Alexa Fluor® 594 Phalloidin (red, panel b) and merged images (panel d).
Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
To demonstrate the phosphorylation of JNK 1 & 2 in a cell based assay, 293 cells were treated with ultraviolet irradiation (UV). Proteins from cell extracts were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were incubated with either 1 µ g/mL ab4821 or 1 µg/mL anti-JNK1 pan. After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar detection method.
To demonstrate the phosphorylation of JNK 1 & 2 in a cell based assay, 293 cells were treated with ultraviolet irradiation (UV). Proteins from cell extracts were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were incubated with either 1 µ g/mL ab4821 or 1 µg/mL anti-JNK1 pan. After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix We
Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
MCF7cells were incubated at 37°C for 4h with vehicle control (0 µM) and different concentrations of cryptotanshinone (ab120666). Increased expression of JNK1+JNK2 (phospho T183 + Y185) in MCF7 cells correlates with an increase in cryptotanshinone concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab4821 at 1/1000 dilution and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.
Immunocytochemistry/ Immunofluorescence - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)This image is courtesy of an Abreview submitted by Mr George Chennell
ab4821 staining JNK1+JNK2 (phospho T183 + Y185) in human foreskin fibroblasts by ICC/IF. The cells were fixed in cytoskeletal fixative, permeabilized in 0.5% Triton X-100 and blocked in 2% dillution buffer (2%BSA + 0.1% Triton X-100) for 1 hour at 25°C. The primary antibody was diluted, 1/100 and incubated with sample for 12 hours. An Alexa Fluor® 594 conjugated goat polyclonal to rabbit IgG, diluted 1/250 was used as secondary.
