Recombinant protein within Human CD19 aa 294-556 / 556.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC
Target Molecular Weight
Predicted band size: 61 kDa
Positive Control
Raji cell lysate, Ramos cell lysate, Daudi cell lysate, Raji, human spleen tissue, Daudi, human tonsils tissue.
Conjugation
unconjugated
Clone Number
JF100-06
Reactivity Data
Tested Verified (internally validated)
Published Reported in literature (not internally validated)
Predicted Predicted reactive (based on sequence homology)
Not recommended Not recommended (failed internal validation)
WB
IF-Cell
IF-Tissue
IHC-P
FC
Human
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
B-lymphocyte antigen CD19, also known as CD19 molecule (Cluster of Differentiation 19), B-Lymphocyte Surface Antigen B4, T-Cell Surface Antigen Leu-12 and CVID3 is a transmembrane protein that in humans is encoded by the gene CD19. In humans, CD19 is expressed in all B lineage cells. Contrary to some early doubts, human plasma cells do express CD19, as confirmed by others. CD19 plays two major roles in human B cells: on the one hand, it acts as an adaptor protein to recruit cytoplasmic signaling proteins to the membrane; on the other, it works within the CD19/CD21 complex to decrease the threshold for B cell receptor signaling pathways. Due to its presence on all B cells, it is a biomarker for B lymphocyte development, lymphoma diagnosis and can be utilized as a target for leukemia immunotherapies.
Background References
1. Walls A et al. Characterization of B-Cells in tonsils of patients diagnosed with pediatric autoimmune neuropsychiatric disorder associated streptococcus. Int J Pediatr Otorhinolaryngol 80:49-52 (2016).
2. Shi H et al. Long non-coding RNA expression profile in minor salivary gland of primary Sj gren\'s syndrome. Arthritis Res Ther 18:109 (2016).
Tissue Specificity
Detected on marginal zone and germinal center B cells in lymph nodes. Detected on blood B cells (at protein level).
Post-translational Modification
Phosphorylated on tyrosine following B-cell activation. Phosphorylated on tyrosine residues by LYN. Tyrosine residues are phosphorylated sequentially after activation of the B cell receptor. Phosphorylation of Tyr-531 is extremely rapid, followed by phosphorylation at Tyr-409. In contrast, phosphorylation of Tyr-500 appears more slowly and is more transient, returning rapidly to basal levels (By similarity).
This data was developed using ET1702-93, the same antibody clone in a different buffer formulation. Western blot analysis of CD19 on different lysates with Rabbit anti-CD19 antibody (ET1702-93) at 1/10,000 dilution.
Lane 1: Raji cell lysate Lane 2: Ramos cell lysate Lane 3: 293T cell lysate (negative) Lane 4: Daudi cell lysate Lane 5: K-562 cell lysate (negative)
Lysates/proteins at 20 µg/Lane.
Predicted band size: 61 kDa Observed band size: 90 kDa
Exposure time: 25 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-93) at 1/10,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Relative expression (RE)
This data was developed using ET1702-93, the same antibody clone in a different buffer formulation. Immunocytochemistry analysis of Raji (positive) and 293T (negative) labeling CD19 with Rabbit anti-CD19 antibody (ET1702-93) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD19 antibody (ET1702-93) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
This data was developed using ET1702-93, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD19 antibody (ET1702-93) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-93) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
This data was developed using ET1702-93, the same antibody clone in a different buffer formulation. Flow cytometric analysis of Daudi cells labeling CD19.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-93, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
This data was developed using ET1702-93, the same antibody clone in a different buffer formulation. Flow cytometric analysis of Raji cells labeling CD19.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-93, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
This data was developed using ET1702-93, the same antibody clone in a different buffer formulation. Application: IF-Tissue
Species: Human
Site: spleen
Sample: Paraffin-embedded section
Antibody concentration: 1/200
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