AB321974

Anti-Rb (phospho S795) 抗体 [EPR28156-6]

Anti-Rb (phospho S795) antibody [EPR28156-6]

20ul selling size
Recombinant
RabMAb
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Anti-Rb (phospho S795) antibody [EPR28156-6] is a Rabbit Monoclonal antibody that is used in Rb (phospho S795) Dot Blot, Western Blot. Suitable for Human samples.

Rb (RB1) is a critical tumor suppressor that prevents transcription of E2F-regulated genes. CDK-dependent phosphorylation of RB1 induces its dissociation from E2Fs, thereby activating transcription of E2F responsive genes and triggering entry into S phase.

別名を表示する

Retinoblastoma-associated protein, p105-Rb, p110-RB1, pRb, pp110, Rb, RB1

4 Images

Supplier Data

Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974)

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

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Rb antibody [EPR17512-291] (ab181616) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) serum starvation for 48h, whole cell lysate (untreated membrane) at 50 µg

Lane 2:

Jurkat serum starvation for 48h, then treated with 10% FBS for 20h, whole cell lysate (untreated membrane) at 50 µg

Lane 3:

Jurkat serum starvation for 48h, then treated with 10% FBS for 20h, 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: 106 kDa,36 kDa

false

Exposure time: 15s

Supplier Data

Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974)

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

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

This blot was developed using a high sensitivity ECL substrate.

The identity of the bands at approximately 40 kDa, 60 kDa and 280 kDa (in lanes 1-2) are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Rb antibody [EPR17512-291] (ab181616) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) serum starvation for 48h, whole cell lysate (untreated membrane) at 50 µg

Lane 2:

MCF7 serum starvation for 48h, then treated with 10% FBS for 20h, whole cell lysate (untreated membrane) at 50 µg

Lane 3:

MCF7 serum starvation for 48h, then treated with 10% FBS for 20h, 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: 106 kDa,36 kDa

true

Exposure time: 92s

Supplier Data

Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974)

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-Rb antibody [EPR17512-291] (ab181616) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974) at 1/1000 dilution

Lane 1:

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

Lane 2:

293T cells transfected with a human Rb (S795A mutation) 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 1/20000 dilution

Observed band size: 106 kDa,124 kDa

false

Exposure time: 1s

Supplier Data

Dot Blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974)

Dot blot analysis of Rb using ab321974 at 1 : 1000 (0.504 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Human Rb (phospho S795) peptide a
Lane2 : Human Rb (phospho S795) peptide b
Lane3 : Human Rb non-phospho peptide
Lane4 : Human Rb (phospho S794) peptide
Lane5 : Mouse Rb (phospho S788) peptide

Exposure time : 180 seconds.

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

All lanes:

Dot Blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974) at 1/1000 dilution

Lane 1:

Human Rb (phospho S795) peptide a

Lane 2:

Human Rb (phospho S795) peptide b

Lane 3:

Human Rb non-phospho peptide

Lane 4:

Human Rb (phospho S794) peptide

Lane 5:

Mouse Rb (phospho S788) 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

Key facts

宿主種

Rabbit

クローン性

Monoclonal

クローン番号

EPR28156-6

アイソタイプ

IgG

キャリアフリー

交差種

Human

アプリケーション

WB, Dot

applications

免疫原

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

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Reactivity data

{ "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"}, "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": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "Mouse": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "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": "", "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": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-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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製品の詳細

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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出荷温度及び保存条件

製品の状態

Liquid

精製方法

Affinity purification Protein A

バッファー組成

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

出荷温度

Blue Ice

短期保存期間

1-2 weeks

短期保存温度

+4°C

長期保存温度

-20°C

分注に関する情報

Upon delivery aliquot

保管に関する情報

Avoid freeze / thaw cycle

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補足情報

This supplementary information is collated from multiple sources and compiled automatically.

Retinoblastoma protein (Rb) also known as pRb is an important regulatory protein with a molecular weight of approximately 110 kDa. It is mainly expressed in the nucleus of cells. Rb functions as a tumor suppressor by controlling the cell cycle progression from G1 to S phase. Rb becomes active when it is dephosphorylated allowing it to bind and inhibit E2F transcription factors consequently preventing the transcription of genes essential for S phase entry.

Biological function summary

Retinoblastoma protein influences cellular proliferation differentiation and apoptosis. It acts within a larger protein complex modulating various cellular responses. When functional Rb halts uncontrolled cell division important for maintaining normal tissue homeostasis. In its phosphorylated state known as phospho-Rb or phospho-Rb E182 it loses its regulatory capabilities which can lead to unrestrained cell cycle progression.

Pathways

Several involve the retinoblastoma protein. One key pathway is the p53 pathway which Rb interacts with to influence cellular outcomes like cell-cycle arrest and apoptosis. Rb cooperates with proteins like p21 to implement these processes. Additionally it is involved in the cyclin D-dependent kinase 4 (CDK4) pathway which modulates its phosphorylation state influencing binding interactions and cell cycle control.

Dysregulation of retinoblastoma protein is commonly associated with cancer particularly retinoblastoma and breast cancer. In retinoblastoma mutations in the Rb gene directly lead to uncontrolled cell proliferation due to the absence of functional Rb protein. Additionally Rb's connection to the E2F family of transcription factors can become disrupted contributing to oncogenesis in other cancers.

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製品プロトコール

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

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

Tumor suppressor that is a key regulator of the G1/S transition of the cell cycle (PubMed : 10499802). The hypophosphorylated form binds transcription regulators of the E2F family, preventing transcription of E2F-responsive genes (PubMed : 10499802). Both physically blocks E2Fs transactivating domain and recruits chromatin-modifying enzymes that actively repress transcription (PubMed : 10499802). Cyclin and CDK-dependent phosphorylation of RB1 induces its dissociation from E2Fs, thereby activating transcription of E2F responsive genes and triggering entry into S phase (PubMed : 10499802). RB1 also promotes the G0-G1 transition upon phosphorylation and activation by CDK3/cyclin-C (PubMed : 15084261). Directly involved in heterochromatin formation by maintaining overall chromatin structure and, in particular, that of constitutive heterochromatin by stabilizing histone methylation. Recruits and targets histone methyltransferases SUV39H1, KMT5B and KMT5C, leading to epigenetic transcriptional repression. Controls histone H4 'Lys-20' trimethylation. Inhibits the intrinsic kinase activity of TAF1. Mediates transcriptional repression by SMARCA4/BRG1 by recruiting a histone deacetylase (HDAC) complex to the c-FOS promoter. In resting neurons, transcription of the c-FOS promoter is inhibited by BRG1-dependent recruitment of a phospho-RB1-HDAC1 repressor complex. Upon calcium influx, RB1 is dephosphorylated by calcineurin, which leads to release of the repressor complex (By similarity).. (Microbial infection) In case of viral infections, interactions with SV40 large T antigen, HPV E7 protein or adenovirus E1A protein induce the disassembly of RB1-E2F1 complex thereby disrupting RB1's activity.

See full target information RB1 phospho S795

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Abcam product promise

当社は、高品質な試薬を通じてお客様の研究を力強くサポートすることをお約束いたします。ご使用いただく各段階で、常にお客様をサポートできる体制を整えております。万が一、製品が期待通りに機能しない場合は、「Abcam Product Promise」による当社保証制度に基づき、安心してご利用いただけます。

保証に関する詳細については利用規約をご確認ください。

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-Rb (phospho S795) 抗体 [EPR28156-6]

Anti-Rb (phospho S795) antibody [EPR28156-6]

Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974)

Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974)

Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974)

Dot Blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974)

Key facts

宿主種

クローン性

クローン番号

アイソタイプ

キャリアフリー

交差種

アプリケーション

免疫原

Reactivity data

製品の詳細

出荷温度及び保存条件

製品の状態

精製方法

バッファー組成

出荷温度

短期保存期間

短期保存温度

長期保存温度

分注に関する情報

保管に関する情報

補足情報

Biological function summary

Pathways

製品プロトコール

ターゲットの情報

Complementary Products

Secondary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Abcam product promise