CD27/TNFRSF7 Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBL1-08927
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for CD27/TNFRSF7 Overexpression Lysate
CD27/TNFRSF7 Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC204252
Accession#: NM_001242
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: RC204252
Accession#: NM_001242
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
26.9 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.
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 CD27/TNFRSF7 Overexpression Lysate
![Western Blot: CD27/TNFRSF7 Overexpression Lysate [NBL1-08927]](https://resources.bio-techne.com/images/products/CD27-Overexpression-Lysate-Adult-Normal-Western-Blot-NBL1-08927-img0002.jpg "Western Blot: CD27/TNFRSF7 Overexpression Lysate [NBL1-08927]")
Western Blot: CD27/TNFRSF7 Overexpression Lysate [NBL1-08927]
Western Blot: CD27 Overexpression Lysate (Adult Normal) [NBL1-08927] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CD27.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: CD27/TNFRSF7
Membrane-bound CD27 is expressed as a disulfide-linked homodimer (3). CD27 binds to the ligand CD70, a transmembrane glycoprotein that is transiently expressed on activated immune cells such as antigen presenting cells (APCs), dendritic cells (DCs), NK cells, B cells, and T cells (1,2,6,7). The receptor-ligand binding interaction leads to NFkappaB and c-Jun pathway activation which promotes immune stimulation and activation and survival of CD4+ T cells, CD8+ T cells, memory T cells, and NK cells (2,6,7). Both CD27 and CD70 are often abnormally expressed or dysregulated on malignant and cancer cells leading to immune evasion and tumor progression (7). CD27 has become a target of interest of immunotherapies for viral infections, autoimmune disease, and cancer (2). Varlilumab, an agonistic CD27 monoclonal antibody (mAB), has entered clinical trials for the treatment of hematological and solid tumor cancers (1,6). Additional clinical trials are in process that combine varlilumab with other immune checkpoint inhibitors like the programmed cell death protein-1 (PD-1) blocking mAb nivolumab (1,2). Initial results are promising, suggesting that targeting CD27, especially in combination with other therapeutics, may be a promising and effective immunotherapy for a variety of pathologies (1,2,6).
References
1. Starzer AM, Berghoff AS. New emerging targets in cancer immunotherapy: CD27 (TNFRSF7). ESMO Open. 2020;4(Suppl 3):e000629. https://doi.org/10.1136/esmoopen-2019-000629
2. Grant EJ, Nussing S, Sant S, Clemens EB, Kedzierska K. The role of CD27 in anti-viral T-cell immunity. Curr Opin Virol. 2017;22:77-88. https://doi.org/10.1016/j.coviro.2016.12.001
3. Buchan SL, Rogel A, Al-Shamkhani A. The immunobiology of CD27 and OX40 and their potential as targets for cancer immunotherapy. Blood. 2018;131(1):39-48. https://10.1182/blood-2017-07-741025
4. Uniprot (P26842)
5. Uniprot (P41272)
6. van de Ven K, Borst J. Targeting the T-cell co-stimulatory CD27/CD70 pathway in cancer immunotherapy: rationale and potential. Immunotherapy. 2015;7(6):655-667. https://doi.org/10.2217/imt.15.32
7. Flieswasser T, Van den Eynde A, Van Audenaerde J, et al. The CD70-CD27 axis in oncology: the new kids on the block. J Exp Clin Cancer Res. 2022;41(1):12. https://doi.org/10.1186/s13046-021-02215-y
Alternate Names
CD27, TNFRSF7
Gene Symbol
CD27
Additional CD27/TNFRSF7 Products
Product Documents for CD27/TNFRSF7 Overexpression Lysate
Product Specific Notices for CD27/TNFRSF7 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.
Loading...
Loading...
Loading...
Loading...