Recombinant Anti-B MyB (phospho T487) antibody [EPR2204Y] - BSA and Azide free (ab232521)
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) - BSA and Azide free
- Suitable for: Flow Cyt (Intra), Dot blot, WB, IHC-P, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-B MyB (phospho T487) antibody [EPR2204Y] - BSA and Azide free
See all B MyB primary antibodies -
Description
Rabbit monoclonal [EPR2204Y] to B MyB (phospho T487) - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), Dot blot, WB, IHC-P, ICC/IFmore 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
ab232521 is the carrier-free version of ab76009.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
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
- PE Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab306203)
- APC Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab306204)
- HRP Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab306205)
- Alexa Fluor® 488 Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab309798)
- Alexa Fluor® 647 Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab310169)
- Alexa Fluor® 594 Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab310602)
- Alexa Fluor® 555 Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab312130)
- Alexa Fluor® 568 Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab312616)
- Anti-B MyB (phospho T487) antibody [EPR2204Y] (ab76009)
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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab232521 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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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Dot blot |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 79 kDa.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Dot blot
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 79 kDa. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76009).
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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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76009).
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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
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76009).
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76009).
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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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76009).
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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
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76009).
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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
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76009).
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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 minutesThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76009).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab232521 has not yet been referenced specifically in any publications.