Collagen I alpha 1 Overexpression Lysate

Novus Biologicals | Catalog # NBP2-11172

Novus Biologicals
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Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Collagen I alpha 1 Overexpression Lysate

Collagen I alpha 1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC206234

Accession#: NM_000088

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

136.5 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 Collagen I alpha 1 Overexpression Lysate

Western Blot: Collagen I Overexpression Lysate (Adult Normal) [NBP2-11172]

Western Blot: Collagen I Overexpression Lysate (Adult Normal) [NBP2-11172] - Validation of overexpression lysate using anti-DDK antibody. Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC206234 using transfection reagent.

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: Collagen I alpha 1

Collagen is an extracellular matrix protein that serves as a scaffold defining the shape and mechanical properties of many tissues and organs including skin, tendon, artery walls, fibrocartilage, bone and teeth. Collagens are highly conserved and are characterized by an uninterrupted "Glycine X Y" triplet repeat that is a necessary part of the triple helical structure. The extensive family of collagens is composed of several chain types, including fibril-forming interstitial collagens (types I, II, III and V) and basement membrane collagens (type IV), each type containing multiple isoforms. Collagen type I (also known as collagen alpha, COL1A1, and alpha-1 type I collagen) is the largest component of fibrillar collagen found in cartilage and connective tissues. It is synthesized by fibroblasts, osteoblasts, and odontoblasts and has a theoretical molecular weight of 138 kDa.

Type I collagen is a fibril-forming collagen found in most connective tissues and is abundant in bone, cornea, dermis and tendon tissue. Collagens are fibrous, extracellular matrix proteins with high tensile strength and are the major components of connective tissue. Several collagens play a role in cell adhesion, responsible for maintaining normal tissue architecture and function. All collagens contain a triple helix domain and frequently show lateral self-association in order to form complex connective tissues. Post-Golgi LH3 trafficking is essential for collagen homeostasis and for the development and function of multiple organs and tissues (1).

The COL1A1 gene encodes the pro-alpha1 chains of type I collagen protein, whose triple helix is comprised of two alpha1 chains and one alpha2 chain. Mutations in the encoding COL1A1 gene are associated with brittle bone disease (Osteogenesis Imperfecta), cortical hyperostosis (Caffey disease) and disorders that affect the connective tissues (Ehlers-Danlos syndrome) (2). Studies have found that HIF-1 transcription regulation of collagen prolyl hydroxylases regulates collagen deposition, promoting cancer cell alignment along collagen fibers, which enhances invasion and metastasis to lymph nodes and lung tissue by breast cancer cells (3).

References

1. Banushi, B., Forneris, F., Straatman-Iwanowska, A., Strange, A., Lyne, A. M., Rogerson, C.,... Gissen, P. (2016). Regulation of post-Golgi LH3 trafficking is essential for collagen homeostasis. Nat Commun, 7, 12111. doi:10.1038/ncomms12111

2. Lu, Y., Zhang, S., Wang, Y., Ren, X., & Han, J. (2019). Molecular mechanisms and clinical manifestations of rare genetic disorders associated with type I collagen. Intractable Rare Dis Res, 8(2), 98-107. doi:10.5582/irdr.2019.01064

3. Gilkes, D. M., Chaturvedi, P., Bajpai, S., Wong, C. C., Wei, H., Pitcairn, S.,... Semenza, G. L. (2013). Collagen prolyl hydroxylases are essential for breast cancer metastasis. Cancer Res, 73(11), 3285-3296. doi:10.1158/0008-5472.Can-12-3963

Alternate Names

COL1A1, OI4

Gene Symbol

COL1A1

Additional Collagen I alpha 1 Products

Product Documents for Collagen I alpha 1 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 Collagen I alpha 1 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 Collagen I alpha 1 Overexpression Lysate

Showing  1 - 4 of 4 FAQs Showing All

  • Q: How long does delivery take for your collagen 1 alpha 1 antibody?

    A: Our most popular collagen I alpha 1 antibody (NB600-408) is usually in stock and ships priority overnight for next business day delivery in the U.S.

  • Q: I am looking for anti collagen 1 antibody, I see your product calls it collagen I alpha 1 is there a difference?

    A: Collagen I alpha 1 is the most common protein for the collagen 1 protein. There are other subunits as well (such as collagen 1 alpha 2…etc.).

  • Q: The predicted molecular weight says 150kDA but the band on the gel appears at 130kDA why is that?

    A: Differences between the predicted and observed MW can be affected by a number of factors including PTMs, relative charges and experimental factors. These cumulative effect of these differences can be quite substantial with larger molecules such as collagen I alpha 1.

  • Q: We are looking for an anti-Collagen antibody for Flow Cytometry (FACS) analysis in Swine sample. Would you please provide some references if there is any available as well?

    A:

    We currently have one antibody for porcine Collagen validated in flow cytometry (catalog # NBP1-94202).

  • Q: How long does delivery take for your collagen 1 alpha 1 antibody?

    A: Our most popular collagen I alpha 1 antibody (NB600-408) is usually in stock and ships priority overnight for next business day delivery in the U.S.

  • Q: I am looking for anti collagen 1 antibody, I see your product calls it collagen I alpha 1 is there a difference?

    A: Collagen I alpha 1 is the most common protein for the collagen 1 protein. There are other subunits as well (such as collagen 1 alpha 2…etc.).

  • Q: The predicted molecular weight says 150kDA but the band on the gel appears at 130kDA why is that?

    A: Differences between the predicted and observed MW can be affected by a number of factors including PTMs, relative charges and experimental factors. These cumulative effect of these differences can be quite substantial with larger molecules such as collagen I alpha 1.

  • Q: We are looking for an anti-Collagen antibody for Flow Cytometry (FACS) analysis in Swine sample. Would you please provide some references if there is any available as well?

    A:

    We currently have one antibody for porcine Collagen validated in flow cytometry (catalog # NBP1-94202).

  • Q: How long does delivery take for your collagen 1 alpha 1 antibody?

    A: Our most popular collagen I alpha 1 antibody (NB600-408) is usually in stock and ships priority overnight for next business day delivery in the U.S.

  • Q: I am looking for anti collagen 1 antibody, I see your product calls it collagen I alpha 1 is there a difference?

    A: Collagen I alpha 1 is the most common protein for the collagen 1 protein. There are other subunits as well (such as collagen 1 alpha 2…etc.).

  • Q: The predicted molecular weight says 150kDA but the band on the gel appears at 130kDA why is that?

    A: Differences between the predicted and observed MW can be affected by a number of factors including PTMs, relative charges and experimental factors. These cumulative effect of these differences can be quite substantial with larger molecules such as collagen I alpha 1.

  • Q: We are looking for an anti-Collagen antibody for Flow Cytometry (FACS) analysis in Swine sample. Would you please provide some references if there is any available as well?

    A:

    We currently have one antibody for porcine Collagen validated in flow cytometry (catalog # NBP1-94202).

  • Q: How long does delivery take for your collagen 1 alpha 1 antibody?

    A: Our most popular collagen I alpha 1 antibody (NB600-408) is usually in stock and ships priority overnight for next business day delivery in the U.S.

  • Q: I am looking for anti collagen 1 antibody, I see your product calls it collagen I alpha 1 is there a difference?

    A: Collagen I alpha 1 is the most common protein for the collagen 1 protein. There are other subunits as well (such as collagen 1 alpha 2…etc.).

  • Q: The predicted molecular weight says 150kDA but the band on the gel appears at 130kDA why is that?

    A: Differences between the predicted and observed MW can be affected by a number of factors including PTMs, relative charges and experimental factors. These cumulative effect of these differences can be quite substantial with larger molecules such as collagen I alpha 1.

  • Q: We are looking for an anti-Collagen antibody for Flow Cytometry (FACS) analysis in Swine sample. Would you please provide some references if there is any available as well?

    A:

    We currently have one antibody for porcine Collagen validated in flow cytometry (catalog # NBP1-94202).

Showing  1 - 4 of 4 FAQs Showing All
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