BCMA/TNFRSF17 Overexpression Lysate

Western Blot: BCMA/TNFRSF17 Overexpression Lysate [NBL1-17151] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for BCMA.

• Reactivity: Hu
• Applications: WB

BCMA/TNFRSF17 Overexpression Lysate Summary

• Description:

  BCMA/TNFRSF17 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC208851
  
  Accession#: NM_001192
  
  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.
• Gene: TNFRSF17

Applications/Dilutions

• Dilutions:
  • Western Blot
• 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.
• Theoretical MW: 20 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.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

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.

Alternate Names for BCMA/TNFRSF17 Overexpression Lysate

• B cell maturation antigen
• B-cell maturation protein
• BCMA
• BCMAtumor necrosis factor receptor superfamily member 17
• BCMB-cell maturation factor
• CD269 antigen
• CD269
• TNFRSF13A
• TNFRSF17
• tumor necrosis factor receptor superfamily, member 17

Background

B cell maturation antigen (BCMA), also known as tumor necrosis factor receptor superfamily member 17 (TNFRSF17), is a type III transmembrane glycoprotein that plays a role in B cell maturation and differentiation into plasma cells and is a therapeutic target for treatment of multiple myeloma (MM) (1,2). BMCA is synthesized as a protein of 184 amino acids (aa) in length with a theoretical molecular weight of 20.2 kDa consisting of an extracellular N-terminus containing 6 cysteine residues, a transmembrane domain, and an intracellular tumor necrosis factor (TRAF) binding domain (1). BCMA is functionally similar to two other TNFR superfamily members, B cell activation factor receptor (BAFF-R) and transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) (1,2). BCMA is primarily expressed on plasmablasts, plasma cells, and late-stage B cells (1,2).

BCMA has two agonistic ligands: BAFF and a proliferation-inducing ligand (APRIL) (1,2). APRIL has higher affinity for BCMA than BAFF and the binding is mediated by CD138/syndeclin-1 (2,3). Activation of BCMA promotes the growth and survival of plasma cells, or MM cells in disease, through several signaling pathways such as NFkappaB, MEK/ERK, AKT, JNK, and p38 (1,2). In MM cells the BCMA activation and downstream signaling cascade functions to upregulate antiapoptotic proteins including Bcl-2, Bcl-xL, and Mcl-1 and protect the cells against therapeutic agents like dexamethasone (2,3).

Given its specific expression on plasma cells but not memory B cells, naive B cells, or hematopoietic stem cells, BCMA has garnered much interest as a therapeutic target for the treatment of MM (1-4). Current BCMA-targeted immunotherapy strategies include antibody-drug conjugates (ADC), chimeric antigen receptor (CAR) T cells, bispecific T cell engager (BiTE), and bispecific/trispecific antibodies (1-4). CAR T cell therapy in particular has demonstrated promising clinical results (2,4). Still, more research needs to be done to improve the efficacy and risk of relapse following CAR T cell therapy and may also include targeting additional antigens in combination with BCMA or utilizing pharmacological agents to increase antigen density (4).

References

1. Yu, B., Jiang, T., & Liu, D. (2020). BCMA-targeted immunotherapy for multiple myeloma. Journal of hematology & oncology, 13(1), 125. https://doi.org/10.1186/s13045-020-00962-7

2. Cho, S. F., Anderson, K. C., & Tai, Y. T. (2018). Targeting B Cell Maturation Antigen (BCMA) in Multiple Myeloma: Potential Uses of BCMA-Based Immunotherapy. Frontiers in immunology, 9, 1821. https://doi.org/10.3389/fimmu.2018.01821

3. Dalla Palma, B., Marchica, V., Catarozzo, M. T., Giuliani, N., & Accardi, F. (2020). Monoclonal and Bispecific Anti-BCMA Antibodies in Multiple Myeloma. Journal of clinical medicine, 9(9), 3022. https://doi.org/10.3390/jcm9093022

4. Mikkilineni, L., & Kochenderfer, J. N. (2021). CAR T cell therapies for patients with multiple myeloma. Nature reviews. Clinical oncology, 18(2), 71-84. https://doi.org/10.1038/s41571-020-0427-6

Limitations

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.

Bioinformatics

• Gene Symbol: TNFRSF17