2B4/CD244/SLAMF4 Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBL1-08924
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for 2B4/CD244/SLAMF4 Overexpression Lysate
2B4/CD244/SLAMF4 Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC206988
Accession#: NM_016382
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: RC206988
Accession#: NM_016382
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
39 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 2B4/CD244/SLAMF4 Overexpression Lysate
![Western Blot: 2B4/CD244/SLAMF4 Overexpression Lysate [NBL1-08924]](https://resources.bio-techne.com/images/products/CD244-Overexpression-Lysate-Adult-Normal-Western-Blot-NBL1-08924-img0002.jpg "Western Blot: 2B4/CD244/SLAMF4 Overexpression Lysate [NBL1-08924]")
Western Blot: 2B4/CD244/SLAMF4 Overexpression Lysate [NBL1-08924]
Western Blot: 2B4/CD244/SLAMF4 Overexpression Lysate [NBL1-08924] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CD244.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: 2B4/CD244/SLAMF4
The 2B4 receptor has both activating and inhibitory signaling properties (1,4). 2B4's ligand is CD48, which is expressed on hematopoietic cells (4, 5). When bound to CD48, subsequent phosphorylation of the ITSMs occurs, followed by recruitment of SH-2-containing adaptor protein (SAP) and Fyn, leading to phosphorylation of PLCgamma and Vav1, and resulting in NK cell activation (4). In the absence of SAP, Src homology region 2-containing protein tyrosine phosphatases (SHP) get recruited and inhibits NK cell activation (4). In the context of infection such as Epstein-Barr virus (EBV), when lacking SAP, 2B4 has been shown to have an inhibitory function via impaired T cell cytotoxicity against EBV-infected cells (6). While 2B4 has a role in eliciting CD8+ T cell response, overexpression of 2B4 can lead to T cell exhaustion in states of chronic infection (6). Studies have suggested that the antibody blockade of the 2B4-CD48 interaction might enhance T cell and NK cell function, however given both the activating and inhibitory properties of the 2B4 receptor, more research needs to be done to prevent unwanted effects (6).
References
1. Buller CW, Mathew PA, Mathew SO. Roles of NK Cell Receptors 2B4 (CD244), CS1 (CD319), and LLT1 (CLEC2D) in Cancer. Cancers (Basel). 2020 Jul 1;12(7):1755. https://doi.org/10.3390/cancers12071755
2. Catakovic K, Klieser E, Neureiter D, Geisberger R. T cell exhaustion: from pathophysiological basics to tumor immunotherapy. Cell Commun Signal. 2017;15(1):1. https://doi.org/10.1186/s12964-016-0160-z
3. Uniprot (Q9BZW8)
4. Chen Y, Lu D, Churov A, Fu R. Research Progress on NK Cell Receptors and Their Signaling Pathways. Mediators Inflamm. 2020 Jul 24;2020:6437057. https://doi.org/10.1155/2020/6437057.
5. Pahima H, Puzzovio PG, Levi-Schaffer F. 2B4 and CD48: A powerful couple of the immune system. Clin Immunol. 2019 Jul;204:64-68. https://doi.org/10.1016/j.clim.2018.10.014.
6. Waggoner SN, Kumar V. Evolving role of 2B4/CD244 in T and NK cell responses during virus infection. Front Immunol. 2012;3:377. https://doi.org/10.3389/fimmu.2012.00377
Alternate Names
CD244, NAIL, NKR2B4, Nmrk, SLAMF4
Gene Symbol
CD244
Additional 2B4/CD244/SLAMF4 Products
Product Documents for 2B4/CD244/SLAMF4 Overexpression Lysate
Product Specific Notices for 2B4/CD244/SLAMF4 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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