Recombinant Anti-MyoD1 antibody [EPR6653-131] (ab133627)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6653-131] to MyoD1
- Suitable for: ChIC/CUT&RUN-seq, WB, ICC/IF, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-MyoD1 antibody [EPR6653-131]
See all MyoD1 primary antibodies -
Description
Rabbit monoclonal [EPR6653-131] to MyoD1 -
Host species
Rabbit -
Tested applications
Suitable for: ChIC/CUT&RUN-seq, WB, ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human MyoD1 aa 50-150.
Database link: P15172 -
Positive control
- IHC-P: Human Rhabdomyosarcoma tissue WB: Human Rhabdomyosarcoma Cell Line Rh30, RD (Human muscle rhabdomyosarcoma) whole cell lysate ICC/IF: RD (Human muscle rhabdomyosarcoma) cell line. ChIC/CUT&RUN-Seq: RD (Human muscle rhabdomyosarcoma) 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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6653-131 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab133627 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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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5µg |
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WB |
1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 34 kDa).
Abcam recommends using milk as the blocking agent. |
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ICC/IF | (1) |
1/100.
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IHC-P | (5) |
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5µg |
WB
1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 34 kDa). Abcam recommends using milk as the blocking agent. |
ICC/IF
1/100. |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
Involved in muscle differentiation (myogenic factor). Induces fibroblasts to differentiate into myoblasts. Activates muscle-specific promoters. Interacts with and is inhibited by the twist protein. This interaction probably involves the basic domains of both proteins. -
Sequence similarities
Contains 1 basic helix-loop-helix (bHLH) domain. -
Post-translational
modificationsAcetylated by a complex containing EP300 and PCAF. The acetylation is essential to activate target genes. Conversely, its deacetylation by SIRT1 inhibits its function.
Ubiquitinated on the N-terminus; which is required for proteasomal degradation. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 4654 Human
- Omim: 159970 Human
- SwissProt: P15172 Human
- Unigene: 181768 Human
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Alternative names
- bHLHc1 antibody
- Class C basic helix-loop-helix protein 1 antibody
- MYF 3 antibody
see all
Images
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 RD (Human muscle rhabdomyosarcoma) cells and 5µg of ab133627 [EPR6653-131]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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Immunohistochemical analysis of MyoD1 in paraffin embedded Human rhabdomyosarcoma tissue, using ab133627 at a dilution of 1/250.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-MyoD1 antibody [EPR6653-131] (ab133627) at 1/1000 dilution
Lane 1 : Rh30 (Human Rhabdomyosarcoma) Whole Cell Lysate at 5 µg
Lane 2 : Rh30 (Human Rhabdomyosarcoma) Whole Cell Lysate at 10 µg
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 4 minutesThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab133627 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
ab133627 detects a band at 45 kDa, while this differs to its predicted molecular weight of 34 kDa, the banding pattern observed is consistent with what has been described in the literature PMID:19352326.
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All lanes : Anti-MyoD1 antibody [EPR6653-131] (ab133627) at 1/1000 dilution
Lane 1 : RD (Human muscle rhabdomyosarcoma) whole cell lysate
Lane 2 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 34 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 37 secondsBlocking and diluting buffer: 5% NFDM/TBST
Negative control: HEK-293, HeLa (PMID: 17028574 )
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Immunocytochemical analysis of 4% paraformaldehyde fixed, 0.1% TritonX-100 permeabilised RD cell line labeling MyoD1 with ab133627 at 1/100 diltuon. ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as a secondary antibody at 1/1000 dilution. Counterstained with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594). Nuclear staining: DAPI.
Confocal image showing nuclear staining in RD cell line
Negative control: HeLa (PMID: 17028574 ) -
ab133627 showing negative staining in Normal heart tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab133627 showing negative staining in Thyroid gland carcinoma tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab133627 showing negative staining in Normal kidney tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab133627 showing negative staining in Skeletal muscle tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (17)
ab133627 has been referenced in 17 publications.
- Chen L et al. CPNE1 regulates myogenesis through the PERK-eIF2α pathway mediated by endoplasmic reticulum stress. Cell Tissue Res 391:545-560 (2023). PubMed: 36525128
- Feng J et al. TGF-β signaling and Creb5 cooperatively regulate Fgf18 to control pharyngeal muscle development. Elife 11:N/A (2022). PubMed: 36542062
- Hsu JY et al. SIX1 reprograms myogenic transcription factors to maintain the rhabdomyosarcoma undifferentiated state. Cell Rep 38:110323 (2022). PubMed: 35108532
- Jin Z et al. Role of skeletal muscle satellite cells in the repair of osteoporotic fractures mediated by β-catenin. J Cachexia Sarcopenia Muscle 13:1403-1417 (2022). PubMed: 35178895
- Sakai H et al. Uhrf1 governs the proliferation and differentiation of muscle satellite cells. iScience 25:103928 (2022). PubMed: 35243267