LAMP-1/CD107a Overexpression Lysate

Name: LAMP-1/CD107a Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-07733

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-07733

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for LAMP-1/CD107a Overexpression Lysate

LAMP-1/CD107a Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC219208

Accession#: NM_005561

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.

TMW

41.9 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 LAMP-1/CD107a Overexpression Lysate

Western Blot: LAMP-1/CD107a Overexpression Lysate [NBP2-07733]

Western Blot: LAMP1 Overexpression Lysate (Adult Normal) [NBP2-07733] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for LAMP1.

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: LAMP-1/CD107a

LAMP-1 (lysosome-associated membrane protein 1), also known as CD107a (cluster of differentiation 107a), is a major component of lysosomal membranes that plays an important role in lysosomal biogenesis, autophagy, and cholesterol metabolism (1). LAMP-1 is a type I transmembrane glycoprotein that is expressed on plasma membranes and the membranes of endosomes, autolysosomes, and lysosomes (1,2). Additionally, LAMP-1/CD107a is a commonly used marker for natural killer (NK) cell degranulation (3). LAMP-1 and another lysosomal-associated membrane protein, LAMP-2, together make up about half of all lysosome membrane proteins (1). Additionally, LAMP-1 has a role in presenting carbohydrate ligands to selectins (2). Human LAMP-1 protein is comprised of 417 amino acids (aa) with a theoretical molecular weight of 44.8 kDa; however, glycosylation can increase the molecular weight upwards of 120 kDa (1, 4). Structurally, LAMP-1 protein contains a large luminal/extracellular domain (29-382 aa), a helical transmembrane domain (383-405 aa), and a short cytoplasmic tail (406-417 aa) (1,2). Additionally, the protein has many N- and O-linked glycosylation sites which helps with stability in the membrane (1,2).

LAMP-1 plays an important role in autophagy-mediated ATP-release during apoptosis where lysosomes containing intracellular ATP migrate to the plasma membrane and, during exocytosis, LAMP-1 is exposed to the cell surface (5). Studies have found that knockdown of LAMP-1 blocks the ATP release from the cell (5). Furthermore, an absence of LAMP-1 and LAMP-2 leads to an accumulation of lysosomal cholesterol (6). Lysosomal membrane dysfunction or defects has also been associated with disease development (6,7). For example, one feature of pancreatitis is autophagy impairment which is caused by lysosomal dysfunction and a corresponding decrease in lysosomal-membrane associated proteins LAMP-1 and LAMP-2 (7).

References

1. Eskelinen E. L. (2006). Roles of LAMP-1 and LAMP-2 in lysosome biogenesis and autophagy. Molecular aspects of medicine, 27(5-6), 495-502. https://doi.org/10.1016/j.mam.2006.08.005

2. Cheng, X. T., Xie, Y. X., Zhou, B., Huang, N., Farfel-Becker, T., & Sheng, Z. H. (2018). Revisiting LAMP1 as a marker for degradative autophagy-lysosomal organelles in the nervous system. Autophagy, 14(8), 1472-1474. https://doi.org/10.1080/15548627.2018.1482147

3. Krzewski, K., & Coligan, J. E. (2012). Human NK cell lytic granules and regulation of their exocytosis. Frontiers in immunology, 3, 335. https://doi.org/10.3389/fimmu.2012.00335

4. Uniprot (P11279)

5. Wang, Y., Martins, I., Ma, Y., Kepp, O., Galluzzi, L., & Kroemer, G. (2013). Autophagy-dependent ATP release from dying cells via lysosomal exocytosis. Autophagy, 9(10), 1624-1625. https://doi.org/10.4161/auto.25873

6. Schwake, M., Schr0der, B., & Saftig, P. (2013). Lysosomal membrane proteins and their central role in physiology. Traffic (Copenhagen, Denmark), 14(7), 739-748. https://doi.org/10.1111/tra.12056

7. Gukovsky, I., Pandol, S. J., Mareninova, O. A., Shalbueva, N., Jia, W., & Gukovskaya, A. S. (2012). Impaired autophagy and organellar dysfunction in pancreatitis. Journal of gastroenterology and hepatology, 27 Suppl 2(Suppl 2), 27-32. https://doi.org/10.1111/j.1440-1746.2011.07004.x

Long Name

Lysosome-associated Membrane Glycoprotein 1

Alternate Names

CD107a, LAMP1

Gene Symbol

LAMP1

Additional LAMP-1/CD107a Products

Product Documents for LAMP-1/CD107a Overexpression Lysate

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COA

SDS

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Product Specific Notices for LAMP-1/CD107a 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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