Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- MA5-14084 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Actin Muscle Monoclonal Antibody (HHF35)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- This antibody reacts with actin from tissue extracts of uterus, ileum, aorta, diaphragm, heart and smooth muscle cells. It recognizes alpha-actin from skeletal, cardiac, and smooth muscle, and reacts with gamma actin from smooth muscle. This antibody will react with tumors from smooth muscle (leiomyomas and leiomyosarcomas) as well as skeletal muscle tumors (rhabdomyomas and rhabdomyosarcomas). Staining of formalin-fixed tissues requires boiling in 1mM EDTA, pH 8.0, followed by cooling at room temperature for 20 min.
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- HHF35
- Vial size
- 500 µL
- Concentration
- Conc. Not Determined
- Storage
- 4° C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Actin Muscle Specific was performed by loading 25 µg of mouse skeletal muscle (Lane 1), Raji (Lane 2) and Hela (Lane 3) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a Actin Muscle Specific monoclonal antibody (Product # MA5-14084) at a dilution of 1:500 (mouse) and 1:100 (Raji and Hela) overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at approx. 42 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on tissue extracts (30 µg lysate) of Mouse Skeletal Muscle (Lane 1), Rat Skeletal Muscle (Lane 2), Mouse Heart (Lane 3), Rat Heart (Lane 4), Mouse Brain (Lane 5) and Rat Brain (Lane 6). The blot was probed with Anti-Actin Muscle Monoclonal Antibody (Product # MA5-14084, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A 42 kDa band corresponding to Actin Muscle was observed across the muscle tissues tested and not in non muscle tissues.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Actin Muscle Specific (green) showing staining in the cytoplasm of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an Actin Muscle Specific monoclonal antibody (Product # MA5-14084) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 100x.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Formalin-fixed, paraffin-embedded human skeletal Muscle stained with Muscle Specific Actin antibody using peroxidase-conjugate and DAB chromogen. Note cytoplasmic staining of muscle cells.