MyoD1 Recombinant Rabbit Monoclonal Antibody [PSH24-48] - BSA and Azide free
Catalog# HA751944
MyoD1 Recombinant Rabbit Monoclonal Antibody [PSH24-48] - BSA and Azide free
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WB
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IF-Cell
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IHC-P
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FC
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IP
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IF-Tissue
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
MyoD1 Recombinant Rabbit Monoclonal Antibody [PSH24-48] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human MyoD1 aa 1-320.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, FC, IP, IF-Tissue
Target Molecular Weight
Predicted band size: 35 kDa
Positive Control
RD (Human malignant embryonal rhabdomyosarcoma cells) cell lysate, C2C12 (Mouse myoblasts) cell lysate, Rat embryo tissue lysate.
Conjugation
unconjugated
Clone Number
PSH24-48
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IF-Cell
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1:500
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IHC-P
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1:200
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FC
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1:1,000
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IP
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1-2μg/sample
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IF-Tissue
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1:200
Target
Function
MyoD, also known as myoblast determination protein 1, is a protein in animals that plays a major role in regulating muscle differentiation. MyoD, which was discovered in the laboratory of Harold M. Weintraub, belongs to a family of proteins known as myogenic regulatory factors (MRFs). These bHLH (basic helix loop helix) transcription factors act sequentially in myogenic differentiation. Vertebrate MRF family members include MyoD1, Myf5, myogenin, and MRF4 (Myf6). In non-vertebrate animals, a single MyoD protein is typically found. MyoD is one of the earliest markers of myogenic commitment. MyoD is expressed at extremely low and essentially undetectable levels in quiescent satellite cells, but expression of MyoD is activated in response to exercise or muscle tissue damage. The effect of MyoD on satellite cells is dose-dependent; high MyoD expression represses cell renewal, promotes terminal differentiation and can induce apoptosis. Although MyoD marks myoblast commitment, muscle development is not dramatically ablated in mouse mutants lacking the MyoD gene. This is likely due to functional redundancy from Myf5 and/or Mrf4. Nevertheless, the combination of MyoD and Myf5 is vital to the success of myogenesis.
Background References
1. Ahmed AA et al. MYOD1 as a prognostic indicator in rhabdomyosarcoma. Pediatr Blood Cancer. 2021 Sep
2. Di Carlo D et al. Biological Role and Clinical Implications of MYOD1L122R Mutation in Rhabdomyosarcoma. Cancers (Basel). 2023 Mar
Subcellular Location
Nucleus.
Synonyms
bHLHc1 antibody
Class C basic helix-loop-helix protein 1 antibody
MYF 3 antibody
Myf-3 antibody
MYF3 antibody
Myoblast determination protein 1 antibody
Myod 1 antibody
MYOD antibody
MYOD1 antibody
MYOD1_HUMAN antibody
ExpandImages
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☑ Relative expression (RE)
Western blot analysis of MyoD1 on different lysates with Rabbit anti-MyoD1 antibody (HA751944) at 1/5,000 dilution.
Lane 1: RD (Human malignant embryonal rhabdomyosarcoma cells) cell lysate (15 µg/Lane)
Lane 2: HEK-293 (Human embryonic kidney cells) cell lysate (15 µg/Lane)
Lane 3: HeLa (Human cervical adenocarcinoma cells) cell lysate (15 µg/Lane)
Lane 4: C2C12 (Mouse myoblasts) cell lysate (15 µg/Lane)
Lane 5: Rat embryo tissue lysate (30 µg/Lane)
Exposure time: 1 minute 2 seconds; ECL: K1801
Negative expression of MyoD1 protein in HEK-293 cells is consistent with the predicted expression pattern.
HeLa is negative control (PMID: 17028574, PMID: 2123467).
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751944, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 35 kDa
Observed band size: 45 kDa -
☑ Relative expression (RE)
Application: Immunocytochemistry (IF-cell)
Species: Human
Sample: RD (Human malignant embryonal rhabdomyosarcoma cells)/HeLa (Human cervix adenocarcinoma epithelial cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA751944, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (M120505-2, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).
Negative control: HeLa (PMID: 17028574, PMID: 2123467). -
Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Embryo
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, citrate buffer (pH 6.0), pressure cooker, 2 minutes.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751944, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Embryo
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, citrate buffer (pH 6.0), pressure cooker, 2 minutes.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751944, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
☑ Relative expression (RE)
Application: Flow Cytometry (Intra)
Species: Human
Sample: RD (Human malignant embryonal rhabdomyosarcoma cells)/HeLa (Human cervix adenocarcinoma epithelial cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA751944 (1/1,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Negative control: HeLa (PMID: 17028574, PMID: 2123467). -
Immunoprecipitation (IP)
MyoD1 was immunoprecipitated in 0.2 mg RD (Human malignant embryonal rhabdomyosarcoma cell) cell lysate with HA751944 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751944 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: RD cell lysate (input)
Lane 2: HA751944 IP in RD cell lysate
Lane 3: Rabbit IgG instead of HA751944 in RD cell lysate
Exposure time: 1 minute
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary dilution: HA751944, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Predicted band size: 35 kDa
Observed band size: 45 kDa -
Application: Immunofluorescence (IF-tissue)
Species: Mouse
Tissue: Embryo
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA751944, 1/200, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"