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AB327426

Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10]

Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10]

  • RabMAb
  • Recombinant
  • 20ul selling size
  • KO Validated
  • 了解详情

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Rabbit Recombinant Monoclonal Cyclin E1 phospho T77 antibody. Suitable for WB, Dot, IP and reacts with Transfected cell lysate - Human, Human, Synthetic peptide - Human samples.

查看别名

CCNE, G1/S-specific cyclin-E1

5 Images
Immunoprecipitation - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (AB327426)
  • IP

Lab

Immunoprecipitation - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (AB327426)

Cyclin E1 was immunoprecipitated from 0.35 mg HCT 116 (human colorectal carcinoma epithelial cell) treated with 2.5mM Thymidine for 16h whole cell lysate with ab327426 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab327426 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HCT 116 (human colorectal carcinoma epithelial cell) treated with 2.5mM Thymidine for 16h whole cell lysate
Lane 2 : ab327426 IP in HCT 116 (human colorectal carcinoma epithelial cell) treated with 2.5mM Thymidine for 16h whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab327426 in HCT 116 treated with 2.5mM Thymidine for 16h whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 180 seconds.

All lanes:

Immunoprecipitation - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (ab327426) at 1/1000 dilution

Lane 1:

HCT 116 (human colorectal carcinoma epithelial cell) treated with 2.5mM Thymidine for 16h whole cell lysate at 10 µg

Lane 2:

ab327426 at 1/30 IP in HCT 116 (human colorectal carcinoma epithelial cell) treated with 2.5mM Thymidine for 16h whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab327426 in HCT 116 treated with 2.5mM Thymidine for 16h whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 35-50 kDa

false

Exposure time: 180s

Western blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (AB327426)
  • WB

Lab

Western blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (AB327426)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (ab327426) at 1/1000 dilution

Lane 1:

Untreated A549 (human lung carcinoma epithelial cell) whole cell lysate (untreated membrane) at 50 µg

Lane 2:

A549 treated with 2.5mM Thymidine for 16h whole cell lysate (untreated membrane) at 50 µg

Lane 3:

A549 treated with 2.5mM Thymidine for 16h whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 35-50 kDa,36 kDa

false

Exposure time: 48s

Western blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (AB327426)
  • WB

Lab

Western blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (AB327426)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab327426 was shown to bind specifically to CCNE1. Target of interest was observed at 35-50 kD in wild-type HCT116 cell lysates (lanes 1-2) with no signal observed at this size in the CCNE1 knockout cell line (lanes 3-4) (lanes 3-4, knockout cell line ab286494).

The identity of the higher MW band at approximately 150 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (ab327426) at 1/1000 dilution

Lane 1:

Untreated HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate (untreated membrane) at 50 µg

Lane 2:

HCT 116 treated with 2.5mM Thymidine for 16h whole cell lysate (untreated membrane) at 50 µg

Lane 3:

CCNE1 knockout HCT116 whole cell lysate (untreated membrane) at 50 µg

Lane 4:

CCNE1 knockout HCT116 treated with 2.5mM Thymidine for 16h) whole cell lysate (untreated membrane) at 50 µg

Lane 5:

Untreated HCT 116 whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Lane 6:

HCT 116 treated with 2.5mM Thymidine for 16h whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Lane 7:

CCNE1 knockout HCT116 whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Lane 8:

CCNE1 knockout HCT116 treated with 2.5mM Thymidine for 16h whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 35-50 kDa,124 kDa

false

Exposure time: 180s

Western blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (AB327426)
  • WB

Lab

Western blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (AB327426)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa) ;Anti-6X His tag antibody [EPR20547]-CHIP Grade (ab213204) (1 : 5000)(35-50KDa).

All lanes:

Western blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (ab327426) at 1/5000 dilution

Lane 1:

293T cells transfected with a human wild-type CCNE1 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human CCNE1 (T77A) expression vector containing a His-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/200000 dilution

Observed band size: 35-50 kDa,36 kDa

false

Exposure time: 15s

Dot Blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (AB327426)
  • Dot

Lab

Dot Blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (AB327426)

Dot blot analysis of Cyclin E1 using ab327426 at 1 : 1000 (0.504 μg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane 1 : CCNE1 (phospho T77) peptide

Lane 2 : CCNE1 (phospho T77) peptide

Lane 3 : CCNE1 non-phospho peptide

Lane 4 : CCNE1 (phospho S73+ T77) peptide

Lane 5 : CCNE1 (phospho S73) peptide

Exposure time : 180 seconds.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Dot Blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (ab327426) at 1/1000 dilution

Lanes 1 - 2:

CCNE1 (phospho T77) peptide

Lane 3:

CCNE1 non-phospho peptide

Lane 4:

CCNE1 (phospho S73+ T77) peptide

Lane 5:

CCNE1 (phospho S73) peptide

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s

关键信息

宿主种属

Rabbit

克隆

Monoclonal

克隆号

EPR28889-10

亚型

IgG

不含载体蛋白

No

反应种属

Human

应用

Dot, WB, IP

applications

免疫原

The exact immunogen used to generate this antibody is proprietary information.

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反应性数据

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/30", "IP-species-notes": "<p></p>", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>" }, "Synthetic peptide - Human": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "<p></p>", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "Transfected cell lysate - Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }
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产品详情

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

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性能和储存信息

形式
Liquid
纯化工艺
Affinity purification Protein A
存储溶液
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
运输条件
Conditional Ambient
推荐的短期储存时间
1-2 weeks
推荐的短期储存条件
+4°C
推荐的长期储存条件
-20°C
分装信息
Upon delivery aliquot
储存信息
Avoid freeze / thaw cycle
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产品实验方案

For this product, it's our understanding that no specific protocols are required. You can visit:
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靶点信息

Essential for the control of the cell cycle at the G1/S (start) transition.
See full target information CCNE1 phospho T77
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Abcam Product Promise

我们致力于为您的研究提供高质量的试剂，为您科研的每一步提供支持。若我们的产品未能达到预期性能，我们向您提供 Abcam Product Promise 保障。
详情请参阅我们的条款与条件。

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