Anti-CREB (phospho S129 + S133)抗体
参阅全部 CREB 一抗
兔多克隆抗体to CREB (phospho S129 + S133)
Rabbit
适用于: ICC/IF, WBmore details
与反应: Mouse, Human
Synthetic peptide corresponding to Human CREB (phospho S129 + S133).
WB: NIH3T3 cell extract with various treatments; A549 cell extract with various treatments; NIH3T3, A549, SK-N-SH, MCF7, and MDA-MB-231 untreated, EGF-treated or PDGF-treated cell lysates. ICC/IF: 70% confluent log phase HeLa cells.
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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: 7.3
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol, 0.1% BSA
PBS is Ca2+ and Mg2+ free
Immunogen affinity purified
Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a spanning peptide corresponding to the cleavage-site to remove antibody that is reactive with non-phosphorylated CREB. The final product is generated by affinity chromatography using a CREB-derived peptide that is phosphorylated at serines 129 and 133.
多克隆
IgG
Abpromise™承诺保证使用ab10564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 | 1/100 - 1/500. | |
| WB | (2) | 1/1000. Detects a band of approximately 43 kDa (predicted molecular weight: 37 kDa). |
Entrez Gene: 1385 Human
Entrez Gene: 12912 Mouse
Omim: 123810 Human
SwissProt: P16220 Human
SwissProt: Q01147 Mouse
Unigene: 516646 Human
Unigene: 453295 Mouse
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Western blot - Anti-CREB (phospho S129 + S133) antibody (ab10564)
All lanes : Anti-CREB (phospho S129 + S133) antibody (ab10564) at 1/1000 dilution
Lane 1 : NIH3T3 cell extract
Lane 2 : NIH3T3 treated with 50 ng/mL PDGF for 15 minutes cell extract
Lane 3 : NIH3T3 treated with 50 ng/mL PDGF for 15 minutes cell extract, incubated with the non-phosphopeptide corresponding to the phosphopeptide immunogen
Lane 4 : NIH3T3 treated with 50 ng/mL PDGF for 15 minutes cell extract, incubated with a generic phosphoserine-containing peptide
Lane 5 : NIH3T3 treated with 50 ng/mL PDGF for 15 minutes cell extract, incubated with the phosphopeptide immunogen
Lane 6 : NIH3T3 treated with 50 ng/mL PDGF for 15 minutes cell extract, treated with lambda phosphatase
Predicted band size: 37 kDa
The membrane was incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and signals were detected using the Pierce SuperSignal™ method.
Western blot - Anti-CREB (phospho S129 + S133) antibody (ab10564)
All lanes : Anti-CREB (phospho S129 + S133) antibody (ab10564) at 0.5 µg/ml
Lane 1 : Human A549 whole cell extract
Lane 2 : Human A549 whole cell extract stimulated with EGF
Lane 3 : Human A549 whole cell extract treated with pervanadate
Lysates/proteins at 20000 cells per lane.
Secondary
All lanes : Anti-mouse, HRP-conjugated
Developed using the ECL technique.
Predicted band size: 37 kDa
Exposure time: 30 seconds
Western blot - Anti-CREB (phospho S129 + S133) antibody (ab10564)
All lanes : Anti-CREB (phospho S129 + S133) antibody (ab10564) at 1/1000 dilution
Lane 1 : NIH/3T3 cell lysate
Lane 2 : NIH/3T3 treated for 10 minutes with 50 ng/mL of PDGF cell lysate
Lane 3 : NIH/3T3 treated for 10 minutes with 200 ng/mL of EGF cell lysate
Lane 4 : A549 cell lysate
Lane 5 : A549 treated for 10 minutes with 200 ng/mL of EGF cell lysate
Lane 6 : A431 cell lysate
Lane 7 : SK-N-SH cell lysate
Lane 8 : MCF7 cell lysate
Lane 9 : MDA-MB-231 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG - HRP at 1/5000 dilution
Developed using the ECL technique.
Predicted band size: 37 kDa
Observed band size: 35,43 kDawhy is the actual band size different from the predicted?
Detection: Chemiluminescence.
Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S129 + S133) antibody (ab10564)
Immunofluorescent analysis of CREB (phospho S129/pS133) was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with CREB (phospho S129/pS133) Rabbit polyclonal Antibody (ab10564) at 2 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody at a dilution of 1/400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin. Panel d is a merged image showing nuclear localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.
