beta-Actin Overexpression Lysate

Novus Biologicals | Catalog # NBP2-10939

Novus Biologicals
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Key Product Details

Species

Human

Applications

Western Blot

Product Summary for beta-Actin Overexpression Lysate

beta-Actin Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC203643

Accession#: NM_001101

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

Theoretical MW

41.6 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for beta-Actin Overexpression Lysate

Western Blot: beta-Actin Overexpression Lysate [NBP2-10939]

Western Blot: beta-Actin Overexpression Lysate [NBP2-10939]

Western Blot: Beta Actin Overexpression Lysate (Adult Normal) [NBP2-10939] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Beta Actin.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: beta-Actin

Actin is a highly conserved protein involved in cell motility, structure, and integrity. Six functional actin isoforms exist and were named based on their electrophoretic mobility and distribution including alpha-cardiac muscle actin (aCAA), alpha-skeletal muscle actin (aSKA), alpha-smooth muscle actin (aSMA), beta-cytoplasmic actin (bCYA), gamma-cytoplasmic actin (gCYA) and gamma-smooth muscle actin (gSMA) (1). The cytoplasmic isoforms, beta-actin and gamma-actin, are nearly identical, differing by only 4 amino acids at the N-terminus with a theoretical molecular weight of 42 kDa (2). Beta-actin is ubiquitously expressed in eukaryotic cells and considered a housekeeping gene which is frequently used as a loading control for assays involving protein detection, such as Western blotting (3,4).

References

1. Vandekerckhove J, Weber K. 1978. At least six different actins are expressed in a higher mammal: an analysis based on the amino acid sequence of the amino-terminal tryptic peptide. J Mol Biol. 126(4):783-802. PMID: 745245

2. Gimona M, Vandekerckhove J, Goethals M, Herzog M, Lando Z, Small JV. (1994) Beta-actin specific monoclonal antibody. Cell Motil Cytoskeleton. 27(2):108-16. PMID: 8162619

3. Holden VI, Lenio S, Kuick R, Ramakrishnan SK, Shah YM, Bachman MA. (2014) Bacterial siderophores that evade or overwhelm Lipocalin 2 induce HIF-1a and pro-inflammatory cytokine secretion in cultured respiratory epithelial cells Infect Immun. 82(9):3826-36 PMID: 24980968

4. Tsai WL, Yeh PH, Tsai CY, Ting CT, Chiu YH, Tao MH, Li WC, Hung SC. (2016) Efficient Programming of Human Mesenchymal Stem Cell Derived Hepatocytes by Epigenetic Regulations. J. Gastroenterol. Hepatol. 32(1):261-269. PMID: 27218433

Alternate Names

ACTB, betaActin

Gene Symbol

ACTB

Additional beta-Actin Products

Product Documents for beta-Actin Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for beta-Actin Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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FAQs for beta-Actin Overexpression Lysate

Showing  1 - 5 of 5 FAQs Showing All

  • Q: Are the beta-Actin antibodies validated in Simple Western?

    A: 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.

  • Q: Do Beta-Actin antibodies come in lyophilized format?

    A: ACTB antibodies like MAB8969 AND MAB8929 come in lyophilized form.

  • Q: 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.

    A: For homogenized tissue, beta-actin and GAPDH are both fine.

  • Q: What is the theoretical molecular weight for Beta-Actin antibodies?

    A: The TMW for beta Actin antibodies is approximately 42 kDa.

  • Q: Why is beta actin used as a loading control in blotting ?

    A: 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.

  • Q: Are the beta-Actin antibodies validated in Simple Western?

    A: 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.

  • Q: Do Beta-Actin antibodies come in lyophilized format?

    A: ACTB antibodies like MAB8969 AND MAB8929 come in lyophilized form.

  • Q: 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.

    A: For homogenized tissue, beta-actin and GAPDH are both fine.

  • Q: What is the theoretical molecular weight for Beta-Actin antibodies?

    A: The TMW for beta Actin antibodies is approximately 42 kDa.

  • Q: Why is beta actin used as a loading control in blotting ?

    A: 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.

  • Q: Are the beta-Actin antibodies validated in Simple Western?

    A: 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.

  • Q: Do Beta-Actin antibodies come in lyophilized format?

    A: ACTB antibodies like MAB8969 AND MAB8929 come in lyophilized form.

  • Q: 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.

    A: For homogenized tissue, beta-actin and GAPDH are both fine.

  • Q: What is the theoretical molecular weight for Beta-Actin antibodies?

    A: The TMW for beta Actin antibodies is approximately 42 kDa.

  • Q: Why is beta actin used as a loading control in blotting ?

    A: 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.

  • Q: Are the beta-Actin antibodies validated in Simple Western?

    A: 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.

  • Q: Do Beta-Actin antibodies come in lyophilized format?

    A: ACTB antibodies like MAB8969 AND MAB8929 come in lyophilized form.

  • Q: 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.

    A: For homogenized tissue, beta-actin and GAPDH are both fine.

  • Q: What is the theoretical molecular weight for Beta-Actin antibodies?

    A: The TMW for beta Actin antibodies is approximately 42 kDa.

  • Q: Why is beta actin used as a loading control in blotting ?

    A: 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.

  • Q: Are the beta-Actin antibodies validated in Simple Western?

    A: 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.

  • Q: Do Beta-Actin antibodies come in lyophilized format?

    A: ACTB antibodies like MAB8969 AND MAB8929 come in lyophilized form.

  • Q: 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.

    A: For homogenized tissue, beta-actin and GAPDH are both fine.

  • Q: What is the theoretical molecular weight for Beta-Actin antibodies?

    A: The TMW for beta Actin antibodies is approximately 42 kDa.

  • Q: Why is beta actin used as a loading control in blotting ?

    A: 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.

Showing  1 - 5 of 5 FAQs Showing All
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