CD19 Antibody, anti-human,
REAlease
^®

Name: CD19 Antibody, anti-human, REAlease®
Availability: InStock

CD19 Antibody, anti-human,
REAlease
^®
Extended ValidationRecombinant

Clone:

REAL106

Type of antibody:

Releasable antibodies, Primary antibodies, Recombinant antibodies

Applications:

FC, MICS, IHC, IF

Alternative names:

B4, CVID3, Leu-12

Flow cytometry applications forCD19 Antibody, anti-human,
REAlease
^®

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Tested:

QC tested

Products used:

130-112-258, 130-112-091, 130-122-650

Protocols:

Cell surface staining protocol using REAlease™ antibodies

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with REAlease CD19 Complexes or with the corresponding REAL Control (left images) as well as with CD20 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide (PI) fluorescence.

MICS (MACSima Imaging Cyclic Staining) applications forCD19 Antibody, anti-human,
REAlease
^®

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-112-258, 130-112-091

Tissue used:

tonsil

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Description:

Immunofluorescence analysis of a human tonsil cryosection fixed with 4% PFA. The section was stained using CD19 Antibody, anti-human, APC (green) for 10 min at room temperature (A). CD19 Antibody, anti-human, APC staining was released using the REAlease Release Reagent (B). Nuclei were visualized with Hoechst™ (blue). Scale bar: 30 µm.

Conjugate:

APC

Recommended dilution:

1:25

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-112-258, 130-112-091

Tissue used:

tonsil

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Description:

Immunofluorescence analysis of a human tonsil cryosection fixed with ice-cold acetone. The section was stained using CD19 Antibody, anti-human, APC (green) for 10 min at room temperature (A). CD19 Antibody, anti-human, APC staining was released using the REAlease Release Reagent (B). Nuclei were visualized with Hoechst™ (blue). Scale bar: 30 µm.

Extended validation for CD19 Antibody, anti-human,
REAlease
^®

Specificity

Knockout validation

To ensure antibody specificity, the target gene is knocked out in a suitable cell line using the CRISPR/Cas9 system and the knockout is confirmed by sequencing of the target locus. The antibody is considered to bind specifically to the intended epitope if no antibody binding to the knockout cells can be detected. The antibody staining is controlled by fluorescence microscopy and/or flow cytometry.

WT

KO

Fluorescence microscopy image of CD19 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD19-PE (REAL106, red) and counterstained with DRAQ5 (blue) as DNA stain.

Fluorescence microscopy image of CD19 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD19-PE (REAL106, red) and counterstained with DRAQ5 (blue) as DNA stain.

Overlay histogram showing flow cytometric analysis of CD19 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD19-PE, clone (REAL106). Flow cytometry was performed with the MACSQuant

^®

Analyzer. Cell debris, dead cells and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Overlay histogram showing flow cytometric analysis of CD19 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD19-PE, clone (REAL106). Flow cytometry was performed with the MACSQuant

^®

Analyzer. Cell debris, dead cells and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Specifications for CD19 Antibody, anti-human,
REAlease
^®

Overview

Clone REAL106 is an antibody fragment derived from the full CD19 antibody molecule. It displays no binding to Fc receptors. The recombinantly engineered antibody fragments are multimerized to form the REAlease Complex to bind markers with high avidity.

Clone REAL106 recognizes the human CD19 antigen, a type I transmembrane glycoprotein of 95 kDa that belongs to the immunglobulin superfamily. CD19 is expressed on B cells throughout most stages of B cell differentiation, though its expression is down-regulated during their terminal differentiation to plasma cells. Expression of CD19 is also found in the majority of B cell–derived malignancies. CD19 is further present on follicular dendritic cells. On B cells, CD19 associates with CD21, CD81, and CD225 (Leu-13) forming a signal transduction complex.

The REAlease Kits consist of the respective fluorochrome-conjugated REAlease Complexes and the REAlease Support Kit for removal of the REAlease Complexes and optional relabeling with different fluorochrome-conjugated REAlease Complexes.

Alternative names

B4, CVID3, Leu-12

Detailed product information

Technical specifications

Clone	REAL106
Clonality	monoclonal
Isotype control	Control Antibody
Host	cell line
Type of antibody	Releasable antibodies, Primary antibodies, Recombinant antibodies
Species	human
Antigen	CD19
Alternative names of antigen	B4, CVID3, Leu-12
Distribution of antigen	B cells, dendritic cells
RRID	AB_2751067, AB_2751457, AB_2751445, AB_2751098, AB_2751079, AB_2784033, AB_2784034, AB_2784032, AB_2751091

Resources for CD19 Antibody, anti-human,
REAlease
^®

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for CD19 Antibody, anti-human,
REAlease
^®

Publications

Nadler, L. M. et al. (1983) B4, a human B lymphocyte-associated antigen expressed on normal, mitogen-activated, and malignant B lymphocytes. J Immunol 131(1): 244-250
Fujimoto, M. et al. (2000) CD19 regulates intrinsic B lymphocyte signal transduction and activation through a novel mechanism of processive amplification. Immunol. Res. 22(2-3): 281-298
Poe, J. C. et al. (2001) CD19, CD21, and CD22: multifaceted response regulators of B lymphocyte signal transduction. Int. Rev. Immunol. 20(6): 739-762
Tedder, T. F. et al. (2002) CD19-CD21 complex regulates an intrinsic Src family kinase amplification loop that links innate immunity with B-lymphocyte intracellular calcium responses. Biochem. Soc. Trans. 30(4): 807-811

Applications

Extended validation