CD34 Antibody, anti-human

Name: CD34 Antibody, anti-human
Availability: InStock

CD34 Antibody, anti-humanExtended Validation

Clone:

AC136

Type of antibody:

Primary antibodies

Isotype:

mouse IgG2aκ, mouse IgG2a

Applications:

FC, MICS, IF, IHC, MC

Alternative names:

gp105-120, Mucosialin, My10

Flow cytometry applications forCD34 Antibody, anti-human

APC
Biotin
FITC
PE
PE-Vio 770
PerCP-Vio 700
VioBlue
VioGreen

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Reported:

20338611, 24999918, 25757087, all publications

Products used:

130-113-738, 130-113-176

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD34 antibodies as well as with CD45 and CD15 antibodies. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. For all other conjugates the FcR Blocking Reagent has been used to avoid Fc receptor–mediated antibody labeling. The cells were analyzed by flow cytometry using the MACSQuant^® Analyzer. A pre-gate of CD45⁺/CD15^– cells was used. 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 tandems.

MICS (MACSima Imaging Cyclic Staining) applications forCD34 Antibody, anti-human

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-113-738, 130-113-176

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-113-738, 130-113-176

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forCD34 Antibody, anti-human

pure

Conjugate:

pure

Reported:

16857897, 22284662, 29261670

Products used:

130-108-040

Extended validation for CD34 Antibody, anti-human

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 AC136
561	++
581	++
563	++
8G12	++
4H11	++
QBEND/10	+
REA1164	++
Cells were incubated with an excess of purified unconjugated CD34 (AC136) 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

Flow cytometric comparison of different clones for CD34. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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.

No fixation

Post fixation

Comparison of staining pattern on non-fixed and fixed cells using CD34 (AC136). 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 (AC136). 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-human

Overview

The monoclonal antibody clone AC136 detects a class III epitope of the CD34 antigen. This epitope is different than the one recognized by the clone used in the CD34 MicroBead Kits.

The CD34 antigen is a single chain transmembrane glycoprotein, expressed on human hematopoitetic stem and progenitor cells, endothelial progenitor cells, vascular endothelial cells, embryonic fibroblasts, and some cells in fetal and adult nervous tissue. The antigen is absent on fully differentiated hematopoietic cells such as normal peripheral blood lymphocytes, monocytes, granulocytes, erythrocytes, and platelets. Clone AC136 has a similar specifity as the CD34 monoclonal antibody clone 8G12 (HPCA-2).

CD34 antibodies can be used for studies of hematopoiesis and nonhematopoiectic stem cells, phenotyping of hematopoietic stem cells, and studies on phenotyping of hematologic malignancies, endothelial cells, and endothelial progenitor cells (EPCs).

Alternative names

gp105-120, Mucosialin, My10

Detailed product information

Technical specifications

Clone	AC136
Clonality	monoclonal
Isotype	mouse IgG2aκ, mouse IgG2a
Isotype control	Isotype Control Antibody, mouse IgG2a
Host	mouse
Type of antibody	Primary antibodies
Species	human
Antigen	CD34
Alternative names of antigen	gp105-120, Mucosialin, My10
Molecular mass of antigen [kDa]	37
Distribution of antigen	stem cells, chondrocytes, dendritic cells, endothelial cells, fibroblasts, mast cells, osteoblasts, cancer stem cells, CNS cells, hematopoietic stem and progenitor cells, bone marrow, liver, dendritic cells, endothelial cells, fibroblasts, osteoblasts, mast cells, stem cells, cancer stem cells, hematopoietic stem and progenitor cells, bone marrow, liver
Entrez Gene ID	947
RRID	AB_2726005, AB_2726281, AB_2726006, AB_2726278, AB_2726003, AB_2726283, AB_2726008, AB_2726284, AB_2726009, AB_2733225, AB_2733226, AB_2726282, AB_2726007, AB_2726279, AB_2726004, AB_2660378, AB_2726280

Resources for CD34 Antibody, anti-human

Certificates

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Reviews for CD34 Antibody, anti-human

Excellent CD34 Antibody from Miltenyi Biotec

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CD34-FITC, human (130-113-178)

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References for CD34 Antibody, anti-human

Publications

De Paepe, M. E. et al. (2011) Alveolar epithelial cell therapy with human cord blood-derived hematopoietic progenitor cells. Am. J. Pathol. 178(3): 1329-1339
Stull, R. A. et al. (2000) Simultaneous flow cytometric analyses of enhanced green and yellow fluorescent Cytometry 40: 126-134
Vets, S. et al. (2012) Lens epithelium-derived growth factor/p75 qualifies as a target for HIV gene therapy in the NSG mouse model. Mol. Ther. 20(5): 908-917
Bility, M. T. et al. (2012) Generation of a humanized mouse model with both human immune system and liver cells to model hepatitis C virus infection and liver immunopathogenesis. Nat. Protoc. 7(9): 1608-1617
Metsuyanim, S. et al. (2009) Expression of stem cell markers in the human fetal kidney. PLoS One 4(68): e6709
Lian, W. et al. (2014)
Varying levels of 6-keto-prostaglandin F1α and thromboxane B2 in serum and endothelialization and hyperplasia in small-diameter grafts seeded with CD34
⁺
bone marrow cells in canines.
Exp Ther Med. 7(5): 1123-1129
Liu, H. et al. (2013) Single-cell clones of liver cancer stem cells have the potential of differentiating into different types of tumor cells. Cell Death Dis 4: e857

Applications

Extended validation