MyoD Antibody (5.8A) - BSA Free

Novus Biologicals | Catalog # NB100-56511

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Predicted:

Porcine (100%), Rat (100%). Backed by our 100% Guarantee.

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Knockdown Validated

Cited:

Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, IF/IHC, Knockdown Validated, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 5.8A

Format

BSA Free
View all MyoD Primary Antibodies »

Product Specifications

Immunogen

The 5.8A antibody was made against recombinant mouse MyoD protein but it also recognizes human (myf3), rat, and cat homologs. The epitope of this antibody was mapped to a region within aa 180-189 of mouse MyoD (NP_002469).

Specificity

In Rh-30, a ~45 kDa band should be observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Description

There is considerable literature published using the MyoD, Clone 5.8A antibody. The original development publication of the MyoD antibody, Clone 5.8A showed that the antibody detected MyoD in rhabdomysosarcomas by IHC (frozen) but not in normal adult tissues (Dias, 1992) or normal fetal skeletal muscle. The 5.8A clone also detected MyoD1 in a subset of Wilms' tumors and one ectomesenchyoma, neoplasms known to contain myogenic elements. These results led to the concept in 1992 that the 5.8A clone may be useful for differentiating rhabdomyosarcomas from other soft tissue malignancies. However, as there has been a myriad of publications since Clone 5.8A was first described, users are encourage to consult the scientific literature citing Clone 5.8A to determine the suitability of the antibody for their model system.

Scientific Data Images for MyoD Antibody (5.8A) - BSA Free

Immunocytochemistry/ Immunofluorescence: MyoD Antibody (5.8A) - BSA Free [NB100-56511]

Immunocytochemistry/ Immunofluorescence: MyoD Antibody (5.8A) - BSA Free [NB100-56511]

Immunocytochemistry/Immunofluorescence: MyoD Antibody (5.8A) [NB100-56511] - Human myoblasts were stained with MyoD1 antibody, diluted 1:200. ICC/IF image submitted by a verified customer review.
Western Blot: MyoD Antibody (5.8A)BSA Free [NB100-56511]

Western Blot: MyoD Antibody (5.8A)BSA Free [NB100-56511]

Western Blot: MyoD Antibody (5.8A) [NB100-56511] - Insuline-like growth factor (IGF1) antagonized the effects of miR-106a-5p on myogenic differentiation in C2C12. All data were collected from C2C12 myotubes 5 d post differentiation. Western-blot analysis of myogenic regulatory factors (MyoD, MyoG, MyHC) in cells; (H) The statistical results of Figure 3G. Data were presented as mean +/- SEM. n = 3 per group. * p < 0.05, ** p < 0.01. Image collected and cropped by CiteAb from the following publication (https://www.mdpi.com/2073-4425/9/7/333/htm) licensed under a CC-BY license. anti-MyoG (catalog# NB100-56510)
Immunocytochemistry/ Immunofluorescence: MyoD Antibody (5.8A) - BSA Free [NB100-56511]

Immunocytochemistry/ Immunofluorescence: MyoD Antibody (5.8A) - BSA Free [NB100-56511]

Immunocytochemistry/Immunofluorescence: MyoD Antibody (5.8A) [NB100-56511] - Cell Lines Tested: mouse skeletal muscle-derived primary cell population Test Sample Preparation: mouse skeletal muscle digested by collagenase type 2 System: Super Sensitive High Resolution Confocal Laser Microscope (LSM880 with Airyscan) Excitation Wavelength: 488nm Emission Filter: 562 nm. Image using the DyLight 488 form of this antibody. ICC/IF image submitted by a verified customer review.
Western Blot: MyoD Antibody (5.8A)BSA Free [NB100-56511]

Western Blot: MyoD Antibody (5.8A)BSA Free [NB100-56511]

Western Blot: MyoD Antibody (5.8A) [NB100-56511] - Analysis for MyoD expression in various small round cell tumor lines using 1 ug/mL anti-MyoD mAb. The antibody only reacts with a band of approx. 45 kDa in the rhabdomyosarcoma cell line (Rh30, lane 1) but was negative against the primitive neuroectodermal (PFSK-1A, lane 2), lymphoma (EB2, lane 3), neuroblastoma (SK-N-SH, lane 4), and Ewing's sarcoma (SJSA-1, lane 5) cell lines.
Western Blot: MyoD Antibody (5.8A)BSA Free [NB100-56511]

Western Blot: MyoD Antibody (5.8A)BSA Free [NB100-56511]

Western Blot: MyoD Antibody (5.8A) [NB100-56511] - MiR-106a-5p inhibited the myogenic differentiation of C2C12 myoblasts. Western blot analyzed for MyoD, MyoG, MyHC proteins 5 d post differentiation. Image collected and cropped by CiteAb from the following publication (https://www.mdpi.com/2073-4425/9/7/333/htm) licensed under a CC-BY license. anti-MyoG (catalog# NB100-56510)
Immunohistochemistry-Frozen: MyoD Antibody (5.8A) - BSA Free [NB100-56511]

Immunohistochemistry-Frozen: MyoD Antibody (5.8A) - BSA Free [NB100-56511]

Immunohistochemistry-Frozen: MyoD Antibody (5.8A) [NB100-56511] - Clone 5.8A antibody in human tissues. A. Rhabdomyosaroma (nuclei are stained), B. Lymphoma (staining is absent)
Immunocytochemistry/ Immunofluorescence: MyoD Antibody (5.8A) - BSA Free [NB100-56511]

Immunocytochemistry/ Immunofluorescence: MyoD Antibody (5.8A) - BSA Free [NB100-56511]

Immunocytochemistry/Immunofluorescence: MyoD Antibody (5.8A) [NB100-56511] - RD cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-MyoD1 (5.8A) NB100-56511 at a 1:200 dilution overnight at 4C and detected with and anti-mouse DyLight 488 (Green) at a 1:500 dilution. Actin was counterstained with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
MyoD Antibody (5.8A) - BSA Free

MyoD (5.8A) in SJCRH30 Human Cell Line.

MyoD (5.8A) was detected in immersion fixed SJCRH30 human Rhabdomyosarcoma cell line with Mouse anti-MyoD (5.8A) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NB100-56511AF488) (green) at 10 µg/mL overnight at 4C. Cells were counterstained with DAPI (dark blue). Cells were imaged using a 100X objective and digitally deconvolved.
MyoD Antibody (5.8A) - BSA Free

Detection of MyoD (5.8A) in SJCRH30 Human Cell Line by Flow Cytometry.

An intracellular stain was performed on SJCRH30 human Rhabdomyosarcoma cell line with Mouse anti- MyoD (5.8A) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NB100-56511AF488, blue histogram) or matched control antibody (orange histogram) at 10 µg/mL for 30 minutes at RT.
MyoD Antibody (5.8A) - BSA Free

Detection of MyoD (5.8A) in SJCRH30 Human Cell Line by Flow Cytometry.

An intracellular stain was performed on SJCRH30 human Rhabdomyosarcoma cell line with Mouse anti- MyoD (5.8A) Protein-G purified Monoclonal Antibody conjugated to FITC (Catalog # NB100-56511F, blue histogram) or matched control antibody (orange histogram) at 5 µg/mL for 30 minutes at RT.

Applications for MyoD Antibody (5.8A) - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

5 ug/ml

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500

Immunoprecipitation

1 ug/ml

Western Blot

1 ug/ml
Application Notes
There is considerable literature published using the MyoD, Clone 5.8 antibody. The original development publication of the MyoD antibody, Clone 5.8A showed that the antibody detected MyoD in rhabdomysosarcomas by IHC (frozen) but not in normal adult tissues (Dias, 1992) or normal fetal skeletal muscle. The 5.8A clone also detected MyoD1 in a subset of Wilms' tumors and one ectomesenchyoma, neoplasms known to contain myogenic elements. These results led to the concept in 1992 that the 5.8A clone may be useful for differentiating rhabdomyosarcomas from other soft tissue malignancies. However, as there has been a myriad of publications since Clone 5.8A was first described, users are encourage to consult the scientific literature citing Clone 5.8A to determine the suitability of the antibody for their model system. Knockdown and IHC-paraffin validation (PMID: 28775895). Use in FLOW reported in scientific literature (Suparamaniam U A et al). This MyoD antibody is validated for Knockout from a verified customer review.

Reviewed Applications

Read 2 reviews rated 4 using NB100-56511 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: MyoD

The Myogenic determination gene (MyoD) was first identified by the virtue of its ability to convert embryonic mouse fibroblast cells to muscle cells. It was subsequently shown that forced expression of MyoD (human homolog is myf 3) gene in a wide variety of normal and neoplastic cells could either convert the cells to muscle cells or activate a set of the otherwise transcriptionally inactive muscle-specific genes in these cells. The regulatory domain of the MyoD gene product lies within a 70 amino acid region and comprises a basic DNA binding motif and a helix-loop-helix (HLH) dimerization motif. Subsequent studies identified three other genes whose products shared sequence homology for the basic HLH domain of MyoD. These are; myf5, myogenin (human homolog is myf4) and myf6 (also known as MRF4 and herculin).

Long Name

Myoblast Determination Protein 1

Alternate Names

bHLHc1, MYF3, MYOD1, PUM

Entrez Gene IDs

4654 (Human); 17927 (Mouse); 337868 (Rat)

Gene Symbol

MYOD1

UniProt

P15172, P10085 (Mouse)

Additional MyoD Products

Product Documents for MyoD Antibody (5.8A) - BSA Free

Certificate of Analysis

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Product Specific Notices for MyoD Antibody (5.8A) - BSA Free

There is considerable literature published using the MyoD, Clone 5.8A antibody. The original development publication of the MyoD antibody, Clone 5.8A showed that the antibody detected MyoD in rhabdomysosarcomas by IHC (frozen) but not in normal adult tissues (Dias, 1992) or normal fetal skeletal muscle. The 5.8A clone also detected MyoD1 in a subset of Wilms' tumors and one ectomesenchyoma, neoplasms known to contain myogenic elements. These results led to the concept in 1992 that the 5.8A clone may be useful for differentiating rhabdomyosarcomas from other soft tissue malignancies. However, as there has been a myriad of publications since Clone 5.8A was first described, users are encourage to consult the scientific literature citing Clone 5.8A to determine the suitability of the antibody for their model system.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for MyoD Antibody (5.8A) - BSA Free

Customer Reviews for MyoD Antibody (5.8A) - BSA Free (2)

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Customer Images

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  • MyoD Antibody (5.8A)
    Name: Pengpeng Bi
    Application: Immunocytochemistry
    Sample Tested: Cell culture
    Species: Human
    Verified Customer | Posted 09/30/2020
    Human myoblasts were stained by primary antibody MyoD1, diluted 1: 200.
    MyoD Antibody (5.8A) - BSA Free NB100-56511
  • MyoD Antibody (5.8A)
    Name: Francesca Pacifici
    Application: Western Blot
    Sample Tested: C2C12 mouse myoblast cell line and Differentiated C2C12 myoblast cells
    Species: Mouse
    Verified Customer | Posted 11/20/2017
    Western Blot: MyoD Antibody (5.8A) [NB100-56511] - Total protein from C2C12 and differentiated C2C12 mouse myoblast cell line, separated on a 4-12% gel by SDS-PAGE. The membrane was probed with anti-MyoD 1:1000 in non-fat milk
    MyoD Antibody (5.8A) - BSA Free NB100-56511

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Protocols

View specific protocols for MyoD Antibody (5.8A) - BSA Free (NB100-56511):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 4% paraformaldehyde to the dish and fix at room temperature for 10 minutes.
2. Remove the paraformaldehyde and wash the cells in PBS.
3. Permeabilize the cells with 0.1% Triton X100 or other suitable detergent for 2 min.
4. Remove the permeabilization buffer and wash three times for 5 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 5 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 5 minutes each.
10. Counter stain DNA with DAPI if required.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

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