Clone:
REA383
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
Mucosialin

Extended validation for CD34 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 clonesOverlap in epitope recognition with REA383
RAM34++
Cells were incubated with an excess of purified unconjugated CD34 (REA383) 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.
View details
Flow cytometric comparison of different clones for CD34. Bone marrow cells from C57BL/6 mice were stained with CD34 antibodies and with a suitable counterstaining. As a control, CD34 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.
View details
Flow cytometric comparison of different clones for CD34. Bone marrow cells from C57BL/6 mice were stained with CD34 antibodies and with a suitable counterstaining. As a control, CD34 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.
View details
Flow cytometric comparison of different clones for CD34. Bone marrow cells from C57BL/6 mice were stained with CD34 antibodies and with a suitable counterstaining. As a control, CD34 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 CD34. Bone marrow cells from C57BL/6 mice were stained with CD34 antibodies and with a suitable counterstaining. As a control, CD34 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
View details
Comparison of staining pattern on non-fixed and fixed cells using CD34 (REA383). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD34 (REA383). 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 CD34 (REA383). 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 CD34 Antibody, anti-mouse, REAfinity™

Overview

Clone REA383 recognizes the mouse CD34 antigen, a highly glycosylated single-pass type I membrane protein also known as mucosialin. CD34 is highly expressed in hematopoietic progenitor cell lines, as well as on endothelial cells, brain, and testis. It is a adhesion molecule with a role in early hematopoiesis by mediating the attachment of stem cells to the bone marrow extracellular matrix or directly to stromal cells. CD34 acts as a scaffold for the attachment of lineage specific glycans, allowing stem cells to bind to lectins expressed by stromal cells or other marrow components.
Additional information: Clone REA383 displays negligible binding to Fc receptors.

Alternative names

Mucosialin

Detailed product information

Technical specifications

CloneREA383
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD34
Alternative names of antigenMucosialin
Molecular mass of antigen [kDa]37
Distribution of antigenendothelial cells, stem cells, hematopoietic stem and progenitor cells, brain, testicle
Entrez Gene ID12490
RRIDAB_2733638

Resources for CD34 Antibody, anti-mouse, REAfinity™

Documents and Protocols

References for CD34 Antibody, anti-mouse, REAfinity™

Publications

  1. Suda, J. et al. (1992) Two types of murine CD34 mRNA generated by alternative splicing. Blood 79(9): 2288-2295
  2. Brown, J. et al. (1991) The gene encoding the stem cell antigen, CD34, is conserved in mouse and expressed in haemopoietic progenitor cell lines, brain, and embryonic fibroblasts. Int. Immunol. 3(2): 175-184
  3. Nakauchi, H. et al. (1999) Further characterization of CD34-low/negative mouse hematopoietic stem cells. Ann. N. Y. Acad. Sci. 872: 57-66

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