CD19 Overexpression Lysate

Name: CD19 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-08913

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-08913

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for CD19 Overexpression Lysate

CD19 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC202922

Accession#: NM_001770

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.

TMW

60.9 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.

Type

Overexpression

Scientific Data Images for CD19 Overexpression Lysate

Western Blot: CD19 Overexpression Lysate [NBL1-08913]

Western Blot: CD19 Overexpression Lysate (Adult Normal) [NBL1-08913] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CD19.

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: CD19

CD19 (Cluster of Differentiation 19), also known as B-lymphocyte surface antigen B4, is a type 1 transmembrane glycoprotein belonging to immunoglobulin (Ig) subfamily that serves as a biomarker for normal and neoplastic B cells (1,2). CD19 is a co-receptor for the B cell receptor (BCR) signaling complex and has a critical role in regulating B cell signaling and immune response (1,2). The CD19 protein contains an extracellular N-terminus containing two C2 Ig-like domains separated by a helical non-Ig domain, a single pass transmembrane domain, and a highly conserved cytoplasmic C-terminal domain (1,2). The human CD19 protein, encoded by the CD19 gene located on chromosome 16p11.2, is 556 amino acids (aa) in length with a calculated theoretical molecular weight (MW) of 61 kDa and an observed molecular weight of 95 kDa (1-3). CD19 associates with other molecules - CD21, CD81, and CD225 - to form the BCR co-complex, also called the CD19 complex, through CD21 binding to the complement C3d complex (1-3). Complement C3d bridges the BCR with the CD19 complex into lipid rafts of the plasma membrane (1-3). CD19 is capable of modulating B cell development through both BCR-dependent and -independent signaling (1-3). Upon BCR activation, the tyrosine residues of CD19's cytoplasmic tail recruits multiple kinases including Lyn, Vav, and PI3K, amplifying BCR-mediated immune signaling and B cell activation (1-3).

Considering the role of CD19 in BCR signaling and its expression in development from pre-B cells through plasma cells, it is understandable that CD19 dysfunction and abnormal expression is associated with numerous B cell malignancies and autoimmune disorders (1-5). CD19 expression is typically observed at relatively normal levels in B cell acute lymphoblastic leukemia (B-ALL) and chronic lymphoblastic leukemia (CLL) but is often reduced other types of lymphoma including diffuse large B cell lymphoma (DLBCL) and follicular lymphoma (FL) (1,2). On the other hand, CD19 expression is typically increased in autoimmune disorders such as systemic sclerosis (SSc) and multiple sclerosis (MS) as modeled by experimental autoimmune encephalomyelitis (EAE) (2). CD19 has become a therapeutic molecular target for the treatment of B cell lymphomas and autoimmune disorders using monoclonal antibodies (mAbs), bi-specific T cell engaging (BiTE) antibodies, and CD19-specific chimeric antigen receptor (CAR) T cells (1,2,4-6). Although anti-CD19 CAR T cell therapy has become the standard for the treatment of B cell malignancies, patients may experience relapse due to resistance mechanisms (6). Strategies to improve efficacy and limit relapse include combination of CAR T cell therapy with immune checkpoint inhibitors like anti-PD-1 (4,6).

References

1. Wang K, Wei G, Liu D. CD19: a biomarker for B cell development, lymphoma diagnosis and therapy. Exp Hematol Oncol. 2012;1(1):36. https://doi.org/10.1186/2162-3619-1-36

2. Li X, Ding Y, Zi M, et al. CD19, from bench to bedside. Immunol Lett. 2017;183:86-95. https://doi.org/10.1016/j.imlet.2017.01.010

3. Wentink MWJ, van Zelm MC, van Dongen JJM, Warnatz K, van der Burg M. Deficiencies in the CD19 complex. Clin Immunol. 2018;195:82-87. https://doi.org/10.1016/j.clim.2018.07.017

4. Frigault MJ, Maus MV. State of the art in CAR T cell therapy for CD19+ B cell malignancies. J Clin Invest. 2020;130(4):1586-1594. https://doi.org/10.1172/JCI129208

5. Penack O, Koenecke C. Complications after CD19+ CAR T-Cell Therapy. Cancers (Basel). 2020;12(11):3445. https://doi.org/10.3390/cancers12113445

6. Bouziana S, Bouzianas D. Anti-CD19 CAR-T cells: Digging in the dark side of the golden therapy. Crit Rev Oncol Hematol. 2021;157:103096. https://doi.org/10.1016/j.critrevonc.2020.103096

Alternate Names

CD19, CVID3, Leu-12

Gene Symbol

CD19

Additional CD19 Products

Product Documents for CD19 Overexpression Lysate

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COA

SDS

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Product Specific Notices for CD19 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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