Anti-CD133 antibody - Stem Cell Marker (ab19898)
Key features and details
- Rabbit polyclonal to CD133 - Stem Cell Marker
- Suitable for: WB
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-CD133 antibody - Stem Cell Marker
See all CD133 primary antibodies -
Description
Rabbit polyclonal to CD133 - Stem Cell Marker -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat
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Immunogen
Synthetic peptide corresponding to Human CD133 (C terminal) conjugated to keyhole limpet haemocyanin.
Database link: O43490 -
Positive control
- WB: Caco 2, HeLa and human embryonic stem cell lysates.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
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Properties
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Form
Liquid -
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-CD133 antibody - Stem Cell Marker (ab19898)All lanes : Anti-CD133 antibody - Stem Cell Marker (ab19898) at 1 µg/ml
Lane 1 : Caco 2 cell lysate
Lane 2 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 97 kDa
Observed band size: 97 kDaLanes 1 - 2: Merged signal (red and green). Green - ab19898 observed at 97 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab19898 was shown to react with CD133 in western blot. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab19898 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-CD133 antibody (ab19898)All lanes : Anti-CD133 antibody - Stem Cell Marker (ab19898) at 1 µg/ml
Lane 1 : MEL-1 (Human embryonic stem cell, male cell line) Whole Cell Lysate (ab27198)
Lane 2 :HT-29 whole cell lysate (ab3952)
Lane 3 :Caco-2 whole cell lysate (ab3950)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 97 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
ab19898 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. -
Western blot - Anti-CD133 antibody (ab19898)All lanes : Anti-CD133 antibody - Stem Cell Marker (ab19898) at 1 µg/ml
Lane 1 : Human Embryonic Stem Cell Lysate
Lane 2 : Human Embryonic Stem Cell Lysate with CD133 peptide (ab20651) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Predicted band size: 97 kDaAnti-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.
