Western blot analysis of human heart whole tissue lysate and with anti-Cardiac Actin antibody. Western blot analysis was performed on 20 µg human heart lysate. The PVDF membrane was blocked with 5% milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-Cardiac Actin mouse monoclonal, AC1-20.4.2 (Cat. No. 690075) was diluted in blocking buffer (antibody concentration 0.05 µg/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Western blot analysis of human heart whole tissue lysate and with anti-Cardiac Actin antibody. Western blot analysis was performed on 20 µg human heart lysate. The PVDF membrane was blocked with 5% milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-Cardiac Actin mouse monoclonal, AC1-20.4.2 (Cat. No. 690075) was diluted in blocking buffer (antibody concentration 0.05 µg/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Key Features
- Mouse monoclonal
- Suitable for IHC and WB
- Reacts with bovine, chicken, human, mouse and rabbit
- Isotype: IgG1
Product description
| Quantity | 200 µl |
|---|---|
| Antibody Type | Monoclonal |
| Host | Mouse |
| Isotype | IgG1 |
| Conjugate | Unconjugated |
| Application | IHC, WB |
| Purification | Affinity chromatography |
| Reactivity | Bovine, Chicken, Human, Mouse, Rabbit |
| Storage | Short term at 2-8°C; long term storage in aliquots at -20°C; avoid freeze/thaw cycles |
| Intended use | Research use only |
| Clone | AC1-20.4.2 |
| Immunogen | Synthetic NH2 terminus decapeptide of cardiac isoform of actin |
| Concentration | 50 µg/ml (10 µg) |
| Formulation | PBS pH 7.4 with 0.09% sodium azide and 0.5% BSA |
| UniprotID | P68034 (Chicken), P68032 (Human), G1STB6 (Rabbit) |
| Synonym | Actin, alpha cardiac muscle 1, Alpha-cardiac actin [Cleaved into: Actin, alpha cardiac muscle 1, intermediate form], ACTC1, ACTC |
Applications
| Tested applications | Tested dilutions |
|---|---|
| Immunohistochemistry (IHC) - frozen | 1:10 (5 µg/ml; include 0.5 M NaCl in all washing buffers to enhance specificity) |
| Immunohistochemistry (IHC) - paraffin | 1:10 (5 µg/ml; microwave treatment recommended) |
| Western Blot (WB) | 1:1,000 (50 ng/ml; include 1 M NaCl in all washing buffers to enhance specificity) |
Background
Ac1 represents an excellent marker for cardiac tissue; it discriminates fetal (cardiac) a-actin from all other actin isoforms. Fetal actin can be localized in regenerating skeletal muscle after injury (in satellite cells) and in veins of the umbelical cord. Mab Ac1-20.4.2 shows no cross reaction with other actin isoforms present in skeletal and smooth muscle, provided that stringent experimental conditions have been applied.
Polypeptide reacting: Specific for fetal (cardiac) isoform of actin.
References/Publications (9)
Publication
Species
Application
Publication
Domke, L and Franke, W. The cell-cell junctions of mammalian testes.., Cell Tissue Res, 375, 451-482, (2019)
Species
bovine
Application
ICC-IF
Publication
Marrocco, V. et al. PKC and PKN in heart disease. J.Mol.Cell.Cardiol. 128, 212-226 (2019)
Species
mouse
Application
WB
Species
mouse
Application
IHC-IF (paraffin)
Species
mouse
Application
WB
Species
human
Application
IHC
Species
human,rat,bovine
Application
IHC (frozen),IHC (paraffin),ICC-IF
Species
chicken
Application
WB,whole mount
Species
chicken
Application
ICC-IF
Species
human
Application
IHC (frozen),IHC (paraffin)
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FAQs
The concentration of unpurified supernatant, ascites, unpurified guinea pig serum and unpurified rabbit serum is not determined.
The concentration of purified antibodies is mentioned on the datasheet.
For prediluted antibodies the concentration may vary from lot to lot. The concentration of these antibodies is not mentioned on the datasheet and can be requested at support@progen.com.
The concentration of purified antibodies is mentioned on the datasheet.
For prediluted antibodies the concentration may vary from lot to lot. The concentration of these antibodies is not mentioned on the datasheet and can be requested at support@progen.com.
Most of our purified mouse antibodies contain 0.5% BSA as stabilizer. If BSA was added to the antibody solution, it is stated in the datasheet.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
Please find the solvent for reconstitution and the amount of liquid for reconstitution on the antibody datasheet. The buffer formulation after reconstitution is also mentioned in the datasheet.
PROGEN offers different formats for mouse monoclonal antibodies. Different antibodies may not be offered in each format, the following are all possible formats:
- Supernatant and supernatant concentrate: This format contains hybridoma cell culture supernatant. The antibody is not purified and the antibody concentration is not determined. The antibody concentration may vary from lot to lot. Therefore we recommend to titrate the optimal concentration for the application used for each new lot.
- Lyophilized, purified: This format contains purified antibody in lyophilized form. The reconstitution of this antibody is described in the datasheet. The buffer composition after reconstitution is also mentioned on the datasheet.
- Liquid, purified: This format contains purified antibody in liquid format. The concentration is mentioned on the datasheet.
- Prediluted, purified: This format contains purified antibody in liquid format. Most antibodies in this format are diluted to be ready-to-use for IHC with standard tissue. But some antibodies of this format need further dilution for IHC. This is mentioned on the datasheet.
Lyophilized antibodies can be stored at 2-8°C until expiration.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.
The expiration date of our antibodies is indicated on the product label.
We offer bulk amounts starting from 1 mg for many of our mouse monoclonal or recombinant antibodies. Please contact us for further information or use the “bulk button” on the product page.
Most of our antibodies contain 0.09% sodium azide as preservative. If a preservative is added, it is mentioned in the datasheet.
- The addition of 1 M NaCl during the first antibody incubation step and the subsequent washing steps in order to eliminate binding to sarcomeric actin is recommended. The epitope of antibody Ac1-20.4.2 on cardiac actin (located at the very end of the N-terminus) differs only by 1 amino acid from the sarcomeric actin.
- To increase stringency further washing steps could be added.
- It is recommended to omit Tween during the antibody incubation step. In our routine protocols we use PBS, supplemented with 1 M NaCl for primary antibody incubation (1 hour) and 3 washing steps (5 min) with PBS/ 1M NaCl.
The optimal antibody dilution for your specific protocol and application needs to be titrated in your lab with your equipment and sample. The optimal dilution may vary between protocols and samples. A good dilution for starting the titration is the dilution mentioned in the datasheet. If the sample needs a specific treatment (eg. Antigen retrieval for IHC on FFPE sections) this should also be mentioned on the antibody datasheet.
PROGEN antibodies are shipped at ambient temperature. The antibodies are stable at ambient temperature for the shipment period. Please store the antibodies as indicated in the datasheet upon arrival.
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