KLRG1 Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBP2-07698
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for KLRG1 Overexpression Lysate
KLRG1 Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC219705
Accession#: NM_005810
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.
Expression Host: HEK293T
Plasmid: RC219705
Accession#: NM_005810
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.
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
21 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.
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 KLRG1 Overexpression Lysate
![Western Blot: KLRG1 Overexpression Lysate [NBP2-07698]](https://resources.bio-techne.com/images/products/KLRG1-Overexpression-Lysate-Adult-Normal-Western-Blot-NBP2-07698-img0001.jpg "Western Blot: KLRG1 Overexpression Lysate [NBP2-07698]")
Western Blot: KLRG1 Overexpression Lysate [NBP2-07698]
Western Blot: KLRG1 Overexpression Lysate (Adult Normal) [NBP2-07698] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for KLRG1.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: KLRG1
KLRG1's primary ligand is E-cadherin, which is expressed on non-neural epithelial tissues, but the KLRG1 receptor also recognizes both N- and R-cadherin expressed in nervous system tissues (1,3,5,6). E-cadherin binding of KLRG1 leads to ITIM triggering and, ultimately, either reduced T cell proliferation or decreased NK cell cytotoxicity (3,5,6). More precisely, upon differentiation of CD8+ T cells, cells switch from signaling through CD28 and instead signal through the inhibitory KLRG1 receptor molecule (3,4). Phosphorylation of KLRG1's ITIM causes recruitment of two phosphatases, SH2-containing inositol polyphosphate 5-phoshate (SHIP-1) and SH-2 containing protein-tyrosine phosphatase 2 (SHP-2) (3,5). The effectors SHIP-1 and SHP-2 degrade PIP3 to PIP2, preventing Akt phosphorylation and inhibiting proliferation (3,5,6). Conversely, when KLRG1 signaling is blocked in highly differentiated T cells, Akt signaling is restored and cell proliferation resumes (3). An increase in highly differentiated T cells is observed in many age-related infections such as meningitis, pneumonia, and influenza, which also correlates with elevated KLRG1 levels (3). This observation suggests that KLRG1 may be a potential immunotherapeutic target, especially for vaccinations which typically have decreased response in aged patients (3).
References
1. Li Y, Hofmann M, Wang Q, Teng L, Chlewicki LK, Pircher H, Mariuzza RA. Structure of natural killer cell receptor KLRG1 bound to E-cadherin reveals basis for MHC-independent missing self recognition. Immunity. 2009 Jul 17;31(1):35-46. http://doi.org/10.1016/j.immuni.2009.04.019
2. Uniprot(Q96E93)
3. Henson SM, Akbar AN. KLRG1--more than a marker for T cell senescence. Age (Dordr). 2009 Dec;31(4):285-91. http://doi.org/10.1007/s11357-009-9100-9.
4. Thimme R, Appay V, Koschella M, Panther E, Roth E, Hislop AD, Rickinson AB, Rowland-Jones SL, Blum HE, Pircher H. Increased expression of the NK cell receptor KLRG1 by virus-specific CD8 T cells during persistent antigen stimulation. J Virol. 2005 Sep;79(18):12112-6.http://doi.org/10.1128/JVI.79.18.12112-12116.2005.
5. Borys SM, Bag AK, Brossay L, Adeegbe DO. The Yin and Yang of Targeting KLRG1+ Tregs and Effector Cells. Front Immunol. 2022 Apr 29;13:894508. http://doi.org10.3389/fimmu.2022.894508.
6. Van den Bossche J, Malissen B, Mantovani A, De Baetselier P, Van Ginderachter JA. Regulation and function of the E-cadherin/catenin complex in cells of the monocyte-macrophage lineage and DCs. Blood. 2012 Feb 16;119(7):1623-33. http://doi.org/10.1182/blood-2011-10-384289.
Long Name
Killer Cell Lectin-like Receptor Subfamily G Member 1
Alternate Names
CLEC15A, MAFA, MAFAL
Gene Symbol
KLRG1
Additional KLRG1 Products
Product Documents for KLRG1 Overexpression Lysate
Product Specific Notices for KLRG1 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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