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CD16a Cell Lysate | FCGR3A cell lysate

Rat CD16a HEK293 Overexpression Lysate

Applications
Western Blot
Synonyms
CD16a; Rat CD16a HEK293 Overexpression Lysate; Rat CD16a/FCGR3A HEK293 Cell Lysate (WB positive control); Fc fragment of IgG; low affinity IIIa; receptor (CD16a); FCGR3A cell lysate
Ordering
For Research Use Only!
Host
HEK293 Cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading buffer).
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Sequence Positions
190aa
Sequence
Met1-Pro200
Applicable Applications for FCGR3A cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Rat
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the rat FCGR3A (XP_008767961.1) extracellular domain (Met 1-Pro 200) was expressed, fused with a polyhistidine tag at the C-terminus.
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for FCGR3A cell lysate
The Fc receptor with low affinity for IgG (FCGR3, or CD16) is encoded by 2 nearly identical genes, FCGR3A and FCGR3B, resulting in tissue-specific expression of alternative membrane-anchored isoforms. FCGR3A, it is also known as CD16a, encodes a transmembrane protein expressed on activated monocytes/macrophages, natural killer (NK) cells, and a subset of T cells. CD16a/FCGR3A is a receptor expressed on NK cells that facilitates antibody dependent cellular cytotoxicity (ADCC) by binding to the Fc portion of various antibodies. CD16a/FCGR3A also has a broader function. CD16a/FCGR3A is directly involved in the lysis of some virus-infected cells and tumor cells by NK cells, independent of antibody binding. Cross-linking of CD16a/FCGR3A on NK cells resulted in increased intracellular Ca2+ levels and a cascade of biochemical events similar to those activated by the T cell receptor. CD16a/FCGR3A on human NK cells is a lysis receptor that mediates the direct killing of some virus infected and tumor cells, independent of antibody ligation.

This Rat CD16a overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD16a protein from the overexpression lysate was verified.
References
David Dornan, et al. Effect of FCGR2A and FCGR3A variants on CLL outcome. Blood. 2010 116: 4212-4222

NCBI and Uniprot Product Information

Molecular Weight
The recombinant rat FCGR3A comprises 190 amino acids and predicts a molecular mass of 21.7kDa.
The apparent molecular mass of the rat FCGR3A is approximately 28-33kDa in SDS-PAGE under reducing conditions due to glycosylation.

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Product Notes

The FCGR3A (Catalog #AAA8117331) is a Cell Lysate produced from HEK293 Cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 190aa. AAA Biotech's CD16a can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the FCGR3A for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Pro20 0. It is sometimes possible for the material contained within the vial of "CD16a, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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