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Recombinant Human CD19 Protein, Atto 647N Conjugate

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Human PBMC CD4+ and CD8+ T cells were either (A) transduced or (B) not transduced (negative control) with a Human CD19 CAR and then cultured for 11 days. Cells were washed twice with 2mL 1x PBS then stained with a viability dye for 15-30 min at room temp in the dark. Human block (1-001-1) was added during the viability stain to block Fc receptors. Cells were washed twice with Flow Cytometry Staining Buffer (FC001) then stained with Human CD4 PE-Cy7-conjugated Antibody and rhCD19 Atto 647N-conjugated protein (ATM9269) for 25 min at room temp in the dark. Staining was performed using our staining membrane proteins protocol.
Human PBMC CD4+ and CD8+ T cells were either (A) transduced or (B) not transduced (negative control) with a Human CD19 CAR and then cultured for 11 days. Cells were washed twice with 2mL 1x PBS then stained with a ...read more
2 μg/lane of Recombinant Human CD19 Fc Chimera Atto 647N (ATM9269) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 77-92 ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Bioactivity

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Recombinant Human CD19 Protein, Atto 647N Conjugate Summary

Additional Information
Fc Chimera
Details of Functionality
Measured by flow cytometry for its ability to bind anti-Human CD19 Monoclonal Antibody conjugated fluorescent beads.
Source
Chinese Hamster Ovary cell line, CHO-derived human CD19 protein
Human CD19
(Glu21-Lys291)
Accession # P15391		IEGRMD Human IgG1
(Pro100-Lys330)
N-terminusC-terminus
Accession #
N-terminal Sequence
Glu21
Structure / Form
Disulfide-linked homodimer, labeled with Atto 647N via amines

Excitation Wavelength: 647 nm
Emission Wavelength: 667 nm
Protein/Peptide Type
Recombinant Proteins
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Bioactivity
Theoretical MW
57 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.
SDS-PAGE
77-92 kDa, under reducing conditions

Packaging, Storage & Formulations

Storage
Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after opening.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human CD19 Protein, Atto 647N Conjugate

  • B4
  • B-lymphocyte antigen CD19
  • B-lymphocyte surface antigen B4
  • CD19 antigen
  • CD19 molecule
  • CD19
  • CVID3
  • Differentiation antigen CD19
  • Leu-12
  • MGC12802
  • T-cell surface antigen Leu-12

Background

CD19 is a 95 kDa transmembrane glycoprotein that plays a central role in B cell activation and humoral immune responses (1, 2). Mature human CD19 consists of a 272 amino acid (aa) extracellular domain (ECD) with two Ig-like domains, a 22 aa transmembrane segment, and a 243 aa cytoplasmic domain (3). Within the ECD, human CD19 shares 57% amino acid sequence identity with mouse and rat CD19. CD19 is expressed throughout B cell development from pre-B cells to mature B cells, and it is commonly used as a B cell lineage marker (1). It is required for the responsiveness of mature B cell to antigen stimulation, germinal center development, and antibody affinity maturation (4, 5). CD19 associates with the B cell antigen receptor (BCR), CD81, CD38, CD21, CD22, and IFITM1/CD225/Leu13, (6-9). These associations enable CD19 to amplify B cell signaling (7, 9-12) and reduce the threshold for antigen stimulation through the BCR (13). CD19 polymorphisms and up-regulation can lead to the development of autoimmunity by promoting autoantibody production (2).
  1. Wang, K. et al. (2012) Exp. Hematol. Oncol. 1:36.
  2. Tedder, T.F. et al. (1997) Immunity 6:107.
  3. Tedder, T.F. and C.M. Isaacs (1989) J. Immunol. 143:712.
  4. Rickert, R.C. et al. (1995) Nature 376:352.
  5. Engel, P. et al. (1995) Immunity 3:39.
  6. Vences-Catalan, F. et al. (2012) J. Immunol. 137:48.
  7. Deaglio, S. et al. (2007) Blood 109:5390.
  8. Bradbury, L. et al. (1992) J. Immunol. 149:2841.
  9. Krop, I. et al. (1996) J. Immunol. 157:48.
  10. O'Rourke, L.M. et al. (1998) Immunity 8:635.
  11. Buhl, A.M. et al. (1997) J. Exp. Med. 186:1897.
  12. Sato, S. et al. (1995) Proc. Natl. Acad. Sci. USA 92:11558.
  13. Carter, R.H. and D.T. Fearon (1992) Science 256:105.

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