CD19 Antibody, anti-mouse, REAfinity™

Name: CD19 Antibody, anti-mouse, REAfinity™
Availability: InStock

CD19 Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA749

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC, 3D-IF

Alternative names:

B4

Flow cytometry applications forCD19 Antibody, anti-mouse, REAfinity™

APC
APC-Vio 770
Biotin
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
Vio Bright B515
Vio Bright R720
VioBlue

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-112-036, 130-111-884

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Splenocytes from BALB/c mice were stained with CD19 antibodies or with the corresponding REA Control antibodies (left images) as well as with CD45R (B220) 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 fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

MICS (MACSima Imaging Cyclic Staining) applications forCD19 Antibody, anti-mouse, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-112-036, 130-111-884

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

3D-Immunofluorescence applications forCD19 Antibody, anti-mouse, REAfinity™

Vio R667

Conjugate:

Vio R667

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

mouse

Products used:

130-131-141

Tissue clearing:

MACS^® Clearing Kit

Compatible applications:

IF, IHC

Description:

3D-immunofluorescence analysis of mouse lymph node fixed with 4% PFA and subjected to whole-mount immunostaining with CD19 Antibody, anti-mouse, Vio R667 (red). The sample was processed and optically cleared using the MACS^® Clearing Kit and imaged with an UltraMicroscope Blaze light sheet instrument. Sample detail image (A), overview image (B), 3D-rendered maximum intensity projection (C) and 3D-rendered video. Scale bars: 100 µm (A), 500 µm (B, C).

Extended validation for CD19 Antibody, anti-mouse, REAfinity™

Specificity

Epitope competition

In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.

Other clones	Overlap in epitope recognition with REA749
6D5	++
1D3	++
MB19-1	++
Cells were incubated with an excess of purified unconjugated CD19 (REA749) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison

Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.

Flow cytometric comparison of different clones for CD19. C57BL6/6 splenocytes were stained with CD19 antibodies and with a suitable counterstaining. As a control, CD19 antibody staining was omitted and cells were measured in the same channels. 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 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD19. C57BL6/6 splenocytes were stained with CD19 antibodies and with a suitable counterstaining. As a control, CD19 antibody staining was omitted and cells were measured in the same channels. 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 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD19. C57BL6/6 splenocytes were stained with CD19 antibodies and with a suitable counterstaining. As a control, CD19 antibody staining was omitted and cells were measured in the same channels. 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 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD19. C57BL6/6 splenocytes were stained with CD19 antibodies and with a suitable counterstaining. As a control, CD19 antibody staining was omitted and cells were measured in the same channels. 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 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD19. C57BL6/6 splenocytes were stained with CD19 antibodies and with a suitable counterstaining. As a control, CD19 antibody staining was omitted and cells were measured in the same channels. 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 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD19. C57BL6/6 splenocytes were stained with CD19 antibodies and with a suitable counterstaining. As a control, CD19 antibody staining was omitted and cells were measured in the same channels. 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 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD19. C57BL6/6 splenocytes were stained with CD19 antibodies and with a suitable counterstaining. As a control, CD19 antibody staining was omitted and cells were measured in the same channels. 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 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Fixation data

To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.

No fixation

Post fixation

Comparison of staining pattern on non-fixed and fixed cells using CD19 (REA749). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Comparison of staining pattern on non-fixed and fixed cells using CD19 (REA749). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD19 Antibody, anti-mouse, REAfinity™

Overview

Clone REA749 recognizes the mouse CD19 antigen, a type I transmembrane glycoprotein expressed on B cells throughout their development from the early pro–B cell through the mature B cell stages. Its expression is down-regulated during terminal differentiation to plasma cells. CD19 associates with complement receptor CD21 and with CD81 on the cell surface of mature B cells, thereby forming a multi-molecular complex which signals synergistically to membrane IgM. It is also expressed on follicular dendritic cells and peritoneal mast cells.

Additional information: Clone REA749 displays negligible binding to Fc receptors.

Alternative names

B4

Detailed product information

Technical specifications

Clone	REA749
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody, human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	mouse
Antigen	CD19
Alternative names of antigen	B4
Molecular mass of antigen [kDa]	58
Distribution of antigen	B cells, dendritic cells
Entrez Gene ID	12478
RRID	AB_2655823, AB_2655824, AB_2655825, AB_2655826, AB_2655827, AB_2655828, AB_2655829, AB_2655830, AB_2655831, AB_2655832, AB_2655833, AB_2655834, AB_2655835, AB_2751063, AB_2751057, AB_2655836, AB_2655837, AB_2921892, AB_2655822

Resources for CD19 Antibody, anti-mouse, REAfinity™

Documents and Protocols

App notes/customer reports

REAfinity™ Recombinant Antibodies - Background-free analysis of mouse TILs

Certificates

Please follow this

link

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

References for CD19 Antibody, anti-mouse, REAfinity™

Publications

Fearon, D. T. (1993) The CD19-CR2-TAPA-1 complex, CD45 and signaling by the antigen receptor of B lymphocytes. Curr. Opin. Immunol. 5(3): 341-348
Tedder, T. F. et al. (1994) The CD19/CD21 signal transduction complex of B lymphocytes. Immunol. Today 15(9): 437-442
Krop, I. et al. (1996) Self-renewal of B-1 lymphocytes is dependent on CD19. Eur. J. Immunol. 26(1): 238-242

Applications

Extended validation