HLA G Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBL1-11592
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for HLA G Overexpression Lysate
HLA G Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC205216
Accession#: NM_002127
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: RC205216
Accession#: NM_002127
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
35.6 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.
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 HLA G Overexpression Lysate
![Western Blot: HLA G Overexpression Lysate [NBL1-11592]](https://resources.bio-techne.com/images/products/HLA-G-Overexpression-Lysate-Adult-Normal-Western-Blot-NBL1-11592-img0002.jpg "Western Blot: HLA G Overexpression Lysate [NBL1-11592]")
Western Blot: HLA G Overexpression Lysate [NBL1-11592]
Western Blot: HLA G Overexpression Lysate (Adult Normal) [NBL1-11592] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for HLA G.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: HLA-G
Discovered in cytotrophoblasts, HLA-G is involved in fetal maternal immune tolerance and some studies have linked its downregulation with severe preeclampsia (3). HLA-G mediated immune suppression works by impeding cell proliferation, differentiation, cytotoxicity, cytokine secretion and chemotaxis; and activation of regulatory cells and MDSCs or M2 type macrophage. HLA-G is constitutively expressed in immune-privileged sites such as pancreatic islets, mesenchymal stem cells (MSCs) and the cornea. Upregulation of HLA-G occurs in pathological states including cancer, allo-transplantations, viral infections, autoimmune and inflammatory diseases (4). In cancer, HLA-G expression is induced by hypoxia and has been correlated with advanced tumor stage, tumor metastasis, and poor therapeutic response and survival. HLA-G is an attractive tumor associated-antigen (TAA) for immunotherapy and is considered a major immune checkpoint (ICP).
References
1. Loustau, M., Anna, F., Drean, R., Lecomte, M., Langlade-Demoyen, P., & Caumartin, J. (2020). HLA-G Neo-Expression on Tumors. Front Immunol, 11:1685. PMID: 32922387
2. Lin A, Yan WH. (2018) Heterogeneity of HLA-G Expression in Cancers: Facing the Challenges. Front Immunol. 9:2164. PMID: 30319626
3. Djurisic S, Hviid TV. (2014) HLA Class Ib Molecules and Immune Cells in Pregnancy and Preeclampsia. Front Immunol. 5:652. PMID: 25566263
4. Lila N, Rouas-Freiss N, Dausset J, Carpentier A, Carosella ED. (2001) Soluble HLA-G protein secreted by allo-specific CD4+ T cells suppresses the allo-proliferative response: a CD4+ T cell regulatory mechanism. Proc Natl Acad Sci U S A. 98(21):12150-12155. PMID: 11572934
Long Name
Major Histocompatibility Complex, Class I, G
Alternate Names
HLA G, HLA-6.0, HLAG, MHC Class I Antigen G, MHC-G
Gene Symbol
HLA-G
Additional HLA-G Products
Product Documents for HLA G Overexpression Lysate
Product Specific Notices for HLA G 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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