Recombinant Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab76009)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2204Y] to B MyB (phospho T487)
- Suitable for: Dot blot, Flow Cyt (Intra), ICC/IF, WB, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-B MyB (phospho T487) antibody [EPR2204Y]
See all B MyB primary antibodies -
Description
Rabbit monoclonal [EPR2204Y] to B MyB (phospho T487) -
Host species
Rabbit -
Tested applications
Suitable for: Dot blot, Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: 293T transfected with human B MyB (WT) expression vector containing a myc-His-tag®, whole cell lysate and HeLa nuclear fraction lysate. ICC/IF: HeLa cells. IHC-P: Human lung and colon cancer tissues. Flow Cyt (Intra): HeLa cells.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2204Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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ChIP Related Products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab76009 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Dot blot |
1/1000.
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Flow Cyt (Intra) |
1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
ICC/IF |
1/1000.
|
|
WB |
1/1000. Predicted molecular weight: 79 kDa.
|
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IHC-P | (1) |
1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
Notes |
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Dot blot
1/1000. |
Flow Cyt (Intra)
1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/1000. |
WB
1/1000. Predicted molecular weight: 79 kDa. |
IHC-P
1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
Transcription factor involved in the regulation of cell survival, proliferation, and differentiation. Transactivates the expression of the CLU gene. -
Sequence similarities
Contains 3 HTH myb-type DNA-binding domains. -
Post-translational
modificationsPhosphorylated by cyclin A/CDK2 during S-phase. Phosphorylation at Thr-520 is probably involved in transcriptional activity. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 4605 Human
- Omim: 601415 Human
- SwissProt: P10244 Human
- Unigene: 179718 Human
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Alternative names
- B-Myb antibody
- BMyB antibody
- MGC15600 antibody
see all
Images
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Immunohistochemistry analysis of paraffin-embedded Human colon cancer tissue sections labelling B MyB (phospho T487) with ab76009 at 1/8000 dilution. The section was incubated with ab76009 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Nuclear staining on Human colon cancer tissue without alkaline phosphatase treatment (Image A); No signal was detected when tissues were treated with alkaline phosphatase (Image B). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells treated with Alkaline Phosphatase overnight (Left) and untreated HeLa cells (Right) labeling B MyB (phospho T487) with ab76009 at 1/1000 dilution (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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All lanes : Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab76009) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) nuclear fraction lysate
Lane 2 : HeLa nuclear fraction lysate treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 79 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 120 secondsBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
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Immunohistochemistry analysis of paraffin-embedded Human lung cancer tissue sections labelling B MyB (phospho T487) with ab76009 at 1/8000 dilution. The section was incubated with ab76009 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Nuclear staining on Human lung cancer tissue without alkaline phosphatase treatment (Image A); No signal was detected when tissues were treated with alkaline phosphatase (Image B). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling B MyB (phospho T487) with primary antibody anti-B MyB (phospho T487) (ab76009) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution. Confocal image showing nuclear staining in HeLa cells and no staining in HeLa cells with Alkaline Phosphatase treatment 37℃ for 1 hour. Anti-alpha Tubulin antibody (DM1A) - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution. The nuclear counter stain is DAPI (blue).
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All lanes : Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab76009) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) nuclear fraction lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) without nuclear fraction lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 79 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 60 secondsBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
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All lanes : Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab76009) at 1/1000 dilution
Lane 1 : 293T (Human embryonic kidney epithelial cell) transfected with empty vector (vector control), containing a myc-His-tag®, whole cell lysate
Lane 2 : 293T transfected with human B MyB (WT) expression vector containing a myc-His-tag®, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 79 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 20 secondsBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
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Dot blot analysis of B MyB (pT487) phospho peptide (lane 1) and B MyB non-phospho peptide (lane 2) with ab76009 at a 1/1000 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/20,000.
Blocking and dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (24)
ab76009 has been referenced in 24 publications.
- Gallo D et al. CCNE1 amplification is synthetic lethal with PKMYT1 kinase inhibition. Nature 604:749-756 (2022). PubMed: 35444283
- Zhong F et al. Downstream Regulatory Network of MYBL2 Mediating Its Oncogenic Role in Melanoma. Front Oncol 12:816070 (2022). PubMed: 35664780
- Orth MF et al. Systematic multi-omics cell line profiling uncovers principles of Ewing sarcoma fusion oncogene-mediated gene regulation. Cell Rep 41:111761 (2022). PubMed: 36476851
- Man N et al. p300 suppresses the transition of myelodysplastic syndromes to acute myeloid leukemia. JCI Insight 6:N/A (2021). PubMed: 34622806
- Xu Q et al. LINC00346 Sponges miR-30c-2-3p to Promote the Development of Lung Adenocarcinoma by Targeting MYBL2 and Regulating CELL CYCLE Signaling Pathway. Front Oncol 11:687208 (2021). PubMed: 34631522