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L-Selectin/CD62L Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-10699

Novus Biologicals, part of Bio-Techne
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NBP2-10699

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for L-Selectin/CD62L Overexpression Lysate

L-Selectin/CD62L Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC216982

Accession#: NM_000655

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

38.6 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 L-Selectin/CD62L Overexpression Lysate

Western Blot: L-Selectin/CD62L Overexpression Lysate [NBP2-10699]

Western Blot: L-Selectin/CD62L Overexpression Lysate [NBP2-10699]

Western Blot: L-selectin Overexpression Lysate (Adult Normal) [NBP2-10699] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for L-selectin.

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: L-Selectin/CD62L

L-selectin, also known as CD62L, is a type I transmembrane glycoprotein that is primarily expressed on leukocytes and has a role in cell adhesion and migration (1,2). L-selectin is closely related to the other family members E- and P-selectin (1,2). Human L-selectin protein is encoded by SELL and is 372 amino acids (aa) in length with a predicted molecular weight (MW) of ~30 kDa (1,2). However, due to glycosylation the observed MW ranges between 65-100 kDa and glycosylation is cell-type dependent (1,2). The L-selectin protein contains an N-terminal C-type lectin domain (CTLD), an epidermal growth factor (EGF)-like domain, two sequence consensus repeat (CSR) domains, a cleavage site, a transmembrane (TM) domain, and a short cytoplasmic tail (1,2).

L-selectin expressed on leukocytes binds to ligands expressed by endothelial cells where it plays a role in lymphocyte homing to secondary lymphoid organs (2-5). L-selectin specifically recognizes and binds to sulfated sialyl-Lewis epitopes of O-linked glycans (2-4). Ligands for L-selectin include glycosylation-dependent cell adhesion molecule-1 (GlyCAM-1), CD34, mucosal vascular addressin cell adhesion molecule-1 (MAdCAM-1), and P-selectin glycoprotein ligand-1 (PSGL-1) (2,4). Elevated levels of selectin ligands on tumor cells are associated with cancer progression and metastasis (3). High levels of L-selectin and soluble L-selectin (sL-selectin) has been implicated in a number of pathologies from viral infection and allergies, to sepsis and multiple sclerosis (2,4,5). For example, L-selectin has been shown to play a role in human immunodeficiency virus (HIV) infection. HIV envelope glycans, such as gp120, binds to L-selectin/CD62L on CD4+ T cells, facilitating viral adhesion (2,5). A disintegrin and metalloproteinase (ADAM)17 is the primary enzyme responsible for L-selectin shedding in leukocytes, which is triggered in response to inflammatory signals (1,2,5). AMAD17 inhibitors block L-selectin shedding and reduce viral release (2,5). Given their role in cancer and other diseases, selectins and their ligands are potential targets for therapeutic intervention (3,5). For instance, murine models have shown that anti-L-selectin antibodies can delay onset of graft versus host disease (5).

References

1. Ivetic A. (2018). A head-to-tail view of L-selectin and its impact on neutrophil behaviour. Cell and Tissue Research, 371(3), 437-453. https://doi.org/10.1007/s00441-017-2774-x

2. Ivetic, A., Hoskins Green, H. L., & Hart, S. J. (2019). L-selectin: a major regulator of leukocyte adhesion, migration and signaling. Frontiers in Immunology, 10, 1068. https://doi.org/10.3389/fimmu.2019.01068

3. Borsig L. (2018). Selectins in cancer immunity. Glycobiology, 28(9), 648-655. https://doi.org/10.1093/glycob/cwx105

4. Kneuer, C., Ehrhardt, C., Radomski, M. W., & Bakowsky, U. (2006). Selectins-potential pharmacological targets?. Drug Discovery Today, 11(21-22), 1034-1040. https://doi.org/10.1016/j.drudis.2006.09.004

5. Segura, J., He, B., Ireland, J., Zou, Z., Shen, T., Roth, G., & Sun, P. D. (2021). The role of L-Selectin in HIV infection. Frontiers in Microbiology, 12, 725741. https://doi.org/10.3389/fmicb.2021.725741

Alternate Names

CD62L, hLHRc, LAM1, LECAM1, Leu-8, LNHR, LSEL, Lyam-1, LYAM1, PLNHR, SELL, TQ1

Gene Symbol

SELL

Additional L-Selectin/CD62L Products

Product Documents for L-Selectin/CD62L Overexpression Lysate

Certificate of Analysis

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

Product Specific Notices for L-Selectin/CD62L 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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