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Anti-A2B5 antibody [105] - Stem Cell Marker (ab53521)

Overview

  • Product nameAnti-A2B5 antibody [105] - Stem Cell MarkerSee all A2B5 primary antibodies ...
  • Description
    Mouse monoclonal [105] to A2B5 - Stem Cell Marker
  • SpecificityThis antibody reacts with a surface antigen on neurons in the retina, brain, spinal cord and dorsal root ganglia, and with all tested human neuroblastoma cell lines. It does not seem to bind with any leukemic cell lines or with bone marrow cells from patients with leukemia, nor with cells from normal bone marrow. It is cytotoxic to neurons in the presence of guinea-pig complement. Protein A binding is positive. It binds with GQ ganglioside on the plasma membrane of neurons of animals of all species tested to date (Eisenbarth, et al., 1979).
  • Tested applicationsFlow Cyt, IHC-Fr, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Human
  • Immunogen

    Embryonic chicken retinal cells.

  • Positive controlType II astrocytes, human neural progenitors.

Properties

Applications

Our Abpromise guarantee covers the use of ab53521 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
Flow Cyt Flow Cyt: Use 1µg for 106 cells.
IHC-Fr IHC-Fr: Use at an assay dependent dilution.
ICC/IF ICC/IF: Use a concentration of 0.5 - 1 µg/ml.

Target

  • RelevanceA2B5 is a cell surface ganglioside epitope expressed in developing thymic epithelial cells, oligodendrocyte progenitors and neuroendocrine cells.
  • Cellular localizationCell surface

Anti-A2B5 antibody [105] - Stem Cell Marker images

  • ICC/IF image of ab53521 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53521, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgM (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).
  • A2B5 antibody [105] - Stem Cell Marker (ab53521) used in immunocytochemical detection on Cor1 neuronal stem cells. Cor1 neuronal stem cells were fixed in formaldehyde, permeabilized, blocked in 1% BSA for 30 mins at RT. ab53521 was incubated at 1/500 for 16 hours in TBS/BSA/azide/0.5% Triton. Secondary Antibody:anti mouse IgM conjugated: to Alexa Fluor® 488 (1/1000).

    See Abreview

  • Overlay histogram showing SH-SY5Y cells stained with ab53521 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53521, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgM (mu chain) (ab97007) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References for Anti-A2B5 antibody [105] - Stem Cell Marker (ab53521)

This product has been referenced in:
  • Chen LP  et al. Regulation of Olig2 during astroglial differentiation in the subventricular zone of a cuprizone-induced demyelination mouse model. Neuroscience 221:96-107 (2012). Read more (PubMed: 22771621) »
  • Paintlia MK  et al. Modulation of peroxisome proliferator-activated receptor-alpha activity by N-acetyl cysteine attenuates inhibition of oligodendrocyte development in lipopolysaccharide stimulated mixed glial cultures. J Neurochem 105:956-70 (2008). Read more (PubMed: 18205750) »

See all 25 Publications for this product

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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Brain)
Specification Brain
Fixative formalin
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH6
Permeabilization No
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Verified customer

Submitted Nov 22 2010

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Cor1 neuronal stem cells)
Specification Cor1 neuronal stem cells
Fixative Formaldehyde
Permeabilization Yes - TBS/BSA/azide/Triton
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Verified customer

Submitted Apr 15 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"