MyoD1 Recombinant Rabbit Monoclonal Antibody [PSH24-48]
cat.: HA751944
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH24-48 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 35 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human MyoD1 aa 1-320. |
| Positive control: | RD (Human malignant embryonal rhabdomyosarcoma cells) cell lysate, C2C12 (Mouse myoblasts) cell lysate, Rat embryo tissue lysate. |
| Subcellular location: | Nucleus. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC IP |
1:5,000 1:500 1:200 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P15172 Human | P10085 Mouse | Q02346 Rat |
| Alternative names: | bHLHc1 Class C basic helix-loop-helix protein 1 MYF 3 Myf-3 MYF3 Myoblast determination protein 1 Myod 1 MYOD MYOD1 MYOD1_HUMAN Myogenic differentiation 1 Myogenic factor 3 Myogenic factor MYF 3 Myogenin D1 PUM |
Images
|
Fig1:
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 |
|
Fig2:
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). |
|
Fig3:
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. |
|
Fig4:
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. |
|
Fig5:
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). |
|
Fig6:
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 Nano-secondary antibody (NBI01H) 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 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.