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AB323846

Anti-Rb (phospho T821 + T826) antibody [EPR28648-126]

  • Recombinant
  • KO Validated
  • 20ul selling size
  • RabMAb
  • What is this?

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Rabbit Recombinant Monoclonal RB phospho T821 + T826 antibody. Suitable for Dot, IP, WB and reacts with Synthetic peptide - Human, Human samples.
3 Images
Immunoprecipitation - Anti-Rb (phospho T821 + T826) antibody [EPR28648-126] (AB323846)
  • IP

Supplier Data

Immunoprecipitation - Anti-Rb (phospho T821 + T826) antibody [EPR28648-126] (AB323846)

Rb (phospho T821 + T826) was immunoprecipitated from 0.35 mg Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) treated with 100ng/ml Calyculin A for 30min whole cell lysate with ab323846 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323846 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Rb (phospho T821 + T826) antibody [EPR28648-126] (ab323846) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) treated with 100ng/ml Calyculin A for 30min whole cell lysate at 10 µg

Lane 2:

ab323846 at 1/30 IP in Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) treated with 100ng/ml Calyculin A for 30min whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323846 in Wild-type HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate at 10 µg

Secondary

All lanes:

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

Observed band size: 106 kDa

false

Exposure time: 15s

Western blot - Anti-Rb (phospho T821 + T826) antibody [EPR28648-126] (AB323846)
  • WB

Supplier Data

Western blot - Anti-Rb (phospho T821 + T826) antibody [EPR28648-126] (AB323846)

Blocking and diluting buffer and concentration : 1% BSA/TBST.

Performed under reducing conditions.

In Western blot, ab323846 was shown to bind specifically to Rb. Target of interest was observed at 106 kDa in Calyculin A treated wild-type HAP1 cell lysates (lane 2) with no signal observed at this size in Calyculin A treated Rb knockout cell line (lane 4).

Lanes 1-4 on the left untreated membrane with alkaline phosphatase; Lanes 1-4 on the right treated membrane with alkaline phosphatase.

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

In Western blot, Anti-Rb antibody [EPR17512] - (ab181616) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Rb (phospho T821 + T826) antibody [EPR28648-126] (ab323846) at 1/1000 dilution

Lane 1:

Untreated Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate (left: untreated membrane with alkaline phosphatase) at 20 µg

Lane 1:

Untreated Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate (right: treated membrane with alkaline phosphatase) at 20 µg

Lane 2:

Wild-type HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (left: untreated membrane with alkaline phosphatase) at 20 µg

Lane 2:

Wild-type HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (right: treated membrane with alkaline phosphatase) at 20 µg

Lane 3:

Untreated Rb knockout HAP1 whole cell lysate (left: untreated membrane with alkaline phosphatase) at 20 µg

Lane 3:

Untreated Rb knockout HAP1 whole cell lysate (right: treated membrane with alkaline phosphatase) at 20 µg

Lane 4:

Rb knockout HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (left: untreated membrane with alkaline phosphatase) at 20 µg

Lane 4:

Rb knockout HAP1 treated with 100ng/ml Calyculin A for 30min whole cell lysate (right: treated membrane with alkaline phosphatase) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/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: 37s

Dot Blot - Anti-Rb (phospho T821 + T826) antibody [EPR28648-126] (AB323846)
  • Dot

Supplier Data

Dot Blot - Anti-Rb (phospho T821 + T826) antibody [EPR28648-126] (AB323846)

Dot blot analysis of Rb (phospho T821 + T826) using ab323846 at 1 : 1000 (0.489 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Rb (phospho T821 + T826) peptide a
Lane2 : Rb (phospho T821 + T826) peptide b
Lane3 : Rb (phospho T826) peptide
Lane4 : Rb (phospho T821) peptide
Lane5 : Rb non-phospho peptide
Lane6 : Rb (phospho S807) peptide
Lane7 : Rb (phospho S811) peptide

Exposure time : 180 seconds.

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

Antibody predominantly detects double phosphorylation of Threonine 821 and 826. Weak binding to single phospho-T821 peptide observed at high concentration.

All lanes:

Dot Blot - Anti-Rb (phospho T821 + T826) antibody [EPR28648-126] (ab323846) at 1/1000 dilution

Lane 1:

Rb (phospho T821 + T826) peptide a

Lane 2:

Rb (phospho T821 + T826) peptide b

Lane 3:

Rb (phospho T826) peptide

Lane 4:

Rb (phospho T821) peptide

Lane 5:

Rb non-phospho peptide

Lane 6:

Rb (phospho S807) peptide

Lane 7:

Rb (phospho S811) peptide

Secondary

All lanes:

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

false

Exposure time: 180s

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR28648-126

Isotype

IgG

Carrier free

No

Reacts with

Human

Applications

Dot, WB, IP

applications

Immunogen

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

Specificity

<table cellspacing="0" style="border-collapse:collapse; width:399px"><tbody><tr><td style="border-bottom:none; border-left:none; border-right:none; border-top:none; height:19px; vertical-align:bottom; white-space:nowrap; width:398px">Antibody predominantly detects double phosphorylation of Threonine 821 and 826&#46; Weak binding to single phospho-T821 peptide observed at high concentration.</td></tr></tbody></table>

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Complementary Products

Primary Antibodies

AB133446

Anti-Rb (phospho T826) antibody [EPR5351]

20ul selling size|RabMAb|Recombinant

Immunoprecipitation - Anti-Rb (phospho T826) antibody [EPR5351] (AB133446)

  • 0
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Primary Antibodies

AB184796

Anti-Rb (phospho S807) antibody [EPR17732]

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Immunocytochemistry/ Immunofluorescence - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

  • 5
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Primary Antibodies

AB181616

Anti-Rb antibody [EPR17512]

20ul selling size|KO Validated|RabMAb|Recombinant|Advanced Validation

Western blot - Anti-Rb antibody [EPR17512] (AB181616)

  • 0
  • 0
Primary Antibodies

AB173289

Anti-Rb (phospho S780) antibody [EPR182(N)]

20ul selling size|RabMAb|Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (AB173289)

  • 5
  • 1
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "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": { "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>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-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>" }, "Mouse": { "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-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": { "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": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-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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Product details

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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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Shipped at conditions
Conditional Ambient
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle
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Product protocols

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

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 pT821 + T826

Alternative Names

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

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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