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Synthetic peptide conjugated to KLH derived from within residues 800 to the C-terminus of Human CD133.
(Peptide available as ab20651.)
Our Abpromise guarantee covers the use of ab19898 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||1/50 - 1/200. PubMed: 23769181|
|IHC-P||1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 97 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
Image courtesy of Human Protein Atlas
ab19898 staining CD133 in human kidney. Paraffin embedded human kidney was incubated with ab19898 (1/150 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6
ab19898 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
Sample: mouse neural stem cells, isolated from the brain of E14 mouse embryo. IF was performed the day after isolation. The antibody was used in concentration 1:200, and was equally effective at the concentration 1:500.
CD133 staining is shown in red; DAPI staining of nuclei is shown in blue.
Anti-CD133 antibody ab19898 detects a band corresponding to the expected size of CD133 in Human Embryonic Stem Cell Lysate using Western Blotting. It is likely that the band runs higher than the predicted MW of CD133 due to glycosylation of CD133. In mouse neural stem cell and mouse embryonic stem cell lysate this ~110 kDa band was not detected but a reproducible band of ~30 kDa was seen. This could represent a cleavage product but it may also be that the ~30 kDa band represents non-specific binding by ab19898. However, ab19898 has been shown to recognise mouse neural stem cells using ICC. Blocking using the immunising peptide removed both of these bands.
ab19898 at 1/200 staining mouse kidney tissue sections by IHC-P. The tissue was paraformaldehyde fixed and blocked with serum. A heat mediated antigen retrieval step was performed and the tissue was then incubated with ab19898 for 16 hours. An Alexa-Fluor ® 488conjugated goat anti-rabbit antibody was used as the secondary.
ab19898 at 1/100 staining mouse kidney tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in formaldehyde buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
Asynchronous KM-H2 cells were pelleted and labeled by indirect immunofluorescence. Cells were stained with ab19898 (1/200) for 30min at 4'C, washed and then stained with goat anti-rabbit alexafluor 488 (1/200). Forward/Side scatter were used to eliminate cellular debris. The accompanying marker was applied such that only 2% of the IgG control was positive. Based on the accompanying image, approximately 48% of cells exhibited positive staining for anti-CD133. This image is taken from an Abreview.
Paraformaldehyde-fixed, paraffin-embedded human glioblastoma multiforme brain tissue stained for CD133 using ab19898 at 1/100 dilution in immunohistochemical analysis.
Formaldehyde-fixed, 0.1% Triton-BSA permeabilized Caco-2 (human colorectal adenocarcinoma cell line) cells stained for CD133 (red) using ab19898 at 1/50 dilution in ICC/IF, followed by Goat Anti-Rabbit IgG (H+L) Cy3. Nuclei staining with Hoechst (blue).
Formaldehyde-fixed, paraffin-embedded rat kidney tissue stained for CD133 using ab19898 at 1/1500 dilution in immunohistochemical analysis, followed by Goat Anti Rabbit IgG Biotin.