Recombinant Human CD19 Protein, Atto 488 Conjugate

Catalog #: ATJ9269 Datasheet / COA / SDS

Fc Chimera

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ATJ9269-020

Detection of CD19 construct on human CART cells

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Recombinant Human CD19 Protein, Atto 488 Conjugate Summary

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Product Specifications

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

Activity

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-terminus		C-terminus

Accession #

P15391

N-terminal Sequence
Analysis

Glu21

Structure / Form

Disulfide-linked homodimer, labeled with Atto 488 via amines

Excitation Wavelength: 501 nm

Emission Wavelength: 523 nm

Predicted Molecular Mass

57 kDa

SDS-PAGE

77-92 kDa, under reducing conditions

Product Datasheets

ATJ9269

Product Datasheet

COA

SDS

ATJ9269

Formulation	Supplied as a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Shipping	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Scientific Data

Flow Cytometry

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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-A) 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 488-conjugated protein (ATJ9269) for 25 min at room temp in the dark. Staining was performed using our staining membrane proteins protocol.

SDS-PAGE

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2 μg/lane of Recombinant Human CD19 Fc Chimera Atto 488 (ATJ9269) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 77-92 kD and 154-184 kDa, respectively.

Reconstitution Calculator

Background: CD19

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).

References

Wang, K. et al. (2012) Exp. Hematol. Oncol. 1:36.
Tedder, T.F. et al. (1997) Immunity 6:107.
Tedder, T.F. and C.M. Isaacs (1989) J. Immunol. 143:712.
Rickert, R.C. et al. (1995) Nature 376:352.
Engel, P. et al. (1995) Immunity 3:39.
Vences-Catalan, F. et al. (2012) J. Immunol. 137:48.
Deaglio, S. et al. (2007) Blood 109:5390.
Bradbury, L. et al. (1992) J. Immunol. 149:2841.
Krop, I. et al. (1996) J. Immunol. 157:48.
O'Rourke, L.M. et al. (1998) Immunity 8:635.
Buhl, A.M. et al. (1997) J. Exp. Med. 186:1897.
Sato, S. et al. (1995) Proc. Natl. Acad. Sci. USA 92:11558.
Carter, R.H. and D.T. Fearon (1992) Science 256:105.

Entrez Gene IDs

930 (Human); 12478 (Mouse); 365367 (Rat); 397669 (Porcine); 607898 (Canine); 102145514 (Cynomolgus Monkey); 100147468 (Equine)

Alternate Names

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