beta-Actin Antibody (1104805) [Unconjugated]

beta -Actin was detected in perfusion fixed paraffin-embedded sections of mouse Stomach using Rat Anti-Mouse beta -Actin, Monoclonal Antibody (Catalog #MAB11702) at 0.25ug/mL at 37° Celsius for 2 minutes.Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog #TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 555 Goat anti-Rat IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cell membrane. Protocol available in COMET™ Panel Builder.​

beta -Actin was detected in immersion fixed paraffin-embedded sections of mouse colon using Rat Anti-Mouse beta -Actin, Monoclonal Antibody (Catalog # MAB11702) at 0.25ug/mL at 37° Celsius for 2 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 555 Goat anti-Rat IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.​​

beta -Actin was detected in immersion fixed paraffin-embedded sections of mouse liver using Rat Anti-Mouse beta -Actin, Monoclonal Antibody (Catalog # MAB11702) at 0.25ug/mL at 37° Celsius for 2 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 555 Goat anti-Rat IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.

beta ‑Actin was detected in perfusion fixed paraffin-embedded sections of mouse heart using Rat Anti-Mouse beta ‑Actin Monoclonal Antibody (Catalog # MAB11702) at 0.05 µg/ml overnight at 4 °C. Tissue was stained using the HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

beta ‑Actin was detected in perfusion fixed paraffin-embedded sections of mouse liver using Rat Anti-Mouse beta ‑Actin Monoclonal Antibody (Catalog # MAB11702) at 0.05 µg/ml overnight at 4 °C. Tissue was stained using the HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

beta ‑Actin was detected in perfusion fixed paraffin-embedded sections of mouse lung using Rat Anti-Mouse beta ‑Actin Monoclonal Antibody (Catalog # MAB11702) at 0.05 µg/ml overnight at 4 °C. Tissue was stained using the HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

beta ‑Actin was detected in immersion fixed 3T3‑L1 mouse embryonic fibroblast adipose-like cell line using Rat Anti-Mouse beta ‑Actin Monoclonal Antibody (Catalog # MAB11702) at 8 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to the cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Western Blot shows lysates of human cerebellum, mouse cerebellum, rat cerebellum, C2C12 mouse myoblast cell line and NIH‑3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 0.5 µg/ml of Rat Anti-Mouse beta ‑Actin Monoclonal Antibody (Catalog # MAB11702) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for beta ‑Actin at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Left: Simple Western lane view shows lysates of C2C12 mouse myoblast cell line, loaded at 0.1 mg/ml. A specific band was detected for beta ‑Actin at approximately 49 kDa (as indicated) using both 10 µg/ml and 50 µg/ml of Rat Anti-Mouse beta ‑Actin Monoclonal Antibody (Catalog # MAB11702) followed by 1:50 dilution of HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005) in Milk-free Antibody Diluent (Catalog # 043-524). This experiment was conducted under reducing conditions and using the 12-230kDa separation system. Right: Simple Western electropherogram showing the same Rat Anti-Mouse beta ‑Actin Monoclonal Antibody (Catalog # MAB11702) tested at 10 µg/ml (blue line) and 50 µg/ml (green line) in the C2C12 mouse myoblast cell line.

• Reactivity: Mu
• Applications: WB, Simple Western, IHC, ICC/IF, mIF
• Clone: 1104805
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated

beta-Actin Antibody (1104805) [Unconjugated] Summary

• Immunogen: Synthetic Peptide
  Accession # P60710
• Specificity: Detects a synthetic peptide specific for mouse beta-actin around amino acid 50 in Direct ELISA.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Rat
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 1-25 ug/mL
  • Immunohistochemistry 0.05-10 ug/mL
  • Multiplex Immunofluorescence 0.25 ug/mL
  • Simple Western 10-50 ug/mL
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Reconstitution Instructions: Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for beta-Actin Antibody (1104805) [Unconjugated]

• ACTB
• actin, beta
• actin, cytoplasmic 1
• B-Actin
• Beta Actin
• beta cytoskeletal actin
• betaActin
• beta-Actin
• BRWS1
• I(2)-Actin
• PS1TP5-binding protein 1
• PS1TP5BP1

Background

Beta actin (ACTB) is a canonical member of the actin family of proteins, with a molecular weight of approximately 42 kDa. Actins are highly conserved proteins that play pivotal roles in various cellular processes, including the maintenance of the cytoskeleton and cell motility, division, and signaling. ACTB is ubiquitously expressed in all eukaryotic cells and is crucial for structural integrity and motility. It is involved in essential biological processes such as cellular trafficking and the maintenance of cell shape and polarity. Dysregulation of ACTB expression or function can lead to notable pathologies, including cancer progression, where it is implicated in tumor cell migration, invasion, and metastasis. Mutations in the ACTB gene are also associated with developmental disorders and conditions like Baraitser-Winter syndrome. Additionally, ACTB plays a critical role in intracellular transport processes and the functioning of the actin cytoskeleton, highlighting its potential as a biomarker for various diseases and a target for therapeutic interventions.

1. Pollard, T. D., & Cooper, J. A. (2009). Actin, a central player in cell shape and movement. Science, 326(5957), 1208-1212. doi: 10.1126/science.1175862.
2. Perrin, B. J., & Ervasti, J. M. (2010). The actin gene family: Function follows isoform. Cytoskeleton, 67(10), 630-634. doi: 10.1002/cm.20475.
3. Jones, S. L., Korobova, F., Svitkina, T. (2019). Origin of the actin cytoskeleton. Nature Reviews Molecular Cell Biology, 20(11), 675-689. doi: 10.1038/s41580-019-0164-5.

Limitations

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.

FAQs for beta-Actin Antibody (MAB11702). (Showing 1 - 5 of 5 FAQ).

1. Why is beta actin used as a loading control in blotting ?
   • Beta actin is a highly-expressed protein found in all cells, and is considered a house keeping protein whose expression is necessary for any cell's proper functioning. For this reason it is used as a loading control to confirm that the same amount of total protein is loaded in each lane of an SDS-PAGE gel so that appropriate comparisons can be made between the actual proteins of interest in different samples.
2. Are the beta-Actin antibodies validated in Simple Western?
   • Yes, we offer a few beta actin antibodies that have been tested in Simple Western: NBP1-47423, NB600-501, NB600-532, NB600-503, and NB100-56874.
3. Do Beta-Actin antibodies come in lyophilized format?
   • ACTB antibodies like MAB8969 AND MAB8929 come in lyophilized form.
4. What is the theoretical molecular weight for Beta-Actin antibodies?
   • The TMW for beta Actin antibodies is approximately 42 kDa.
5. I wanted to know which of the two housekeeping genes B-actin or GAPDH can be used for best results. I intend to do an experiment to determine expression of IL-6 and TNF-alpha in liver and kidney tissues.
   • For homogenized tissue, beta-actin and GAPDH are both fine.