Antibodies, Proteins, Kits and Reagents for Life Science

311 587 ₸ Product size

100 µl 311 587 ₸

Order now and get it on Wednesday March 10, 2021

Anti-CD133 antibody [EPR20980-45] (ab226355)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20980-45] to CD133
Suitable for: WB, IHC-P
Reacts with: Human

Product name

Anti-CD133 antibody [EPR20980-45]
See all CD133 primary antibodies
Description

Rabbit monoclonal [EPR20980-45] to CD133
Host species

Rabbit
Tested Applications & Species

Application Species
IHC-P
Human

WB
Human

See all applications and species data
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Caco-2, NT2/D1 and HT-29 whole cell lysate. Human pancreas lysate and fetal kidney lysate. IHC-P: Human pancreas tissue. Human pancreatic cancer tissue.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Application	Species
IHC-P	Human
WB	Human

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20980-45
Isotype

IgG
Research areas

Western blot - Anti-CD133 antibody [EPR20980-45] (ab226355)

All lanes : Anti-CD133 antibody [EPR20980-45] (ab226355) at 1/1000 dilution

Lane 1 : Wild-type HAP1 cell lysate at 40 µg
Lane 2 : PROM1 knockout HAP1 cell lysate at 40 µg
Lane 3 : Caco-2 cell lysate at 20 µg
Lane 4 : HeLa cell lysate at 20 µg

Performed under reducing conditions.

Predicted band size: 97 kDa
Observed band size: 110 kDa
why is the actual band size different from the predicted?

Exposure time: 20 minutes

ab226355 was shown to react with CD133 in wild-type HAP1 cells in western blot. Loss of signal was observed when PROM1 knockout sample was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab226355 overnight at 4°C at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1/5000 for 1 hour at room temperature before development with ECL reagent and imaging.
Western blot - Anti-CD133 antibody [EPR20980-45] (ab226355)

All lanes : Anti-CD133 antibody [EPR20980-45] (ab226355) at 1/1000 dilution

Lane 1 : Caco-2 (human colorectal adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : HT-29 (human colorectal adenocarcinoma epithelial cell), whole cell lysate
Lane 3 : NCCIT (human pluripotent embryonic carcinoma epithelial cell), whole cell lysate
Lane 4 : HEK-293T (human embryonic kidney epithelial cell), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 97 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking: 5% NFDM/TBST.

The MW observed is consistent with the literature (PMID 20717901; PMID 18645205).

Negative control: HEK-293T (PMID 20167130).
Western blot - Anti-CD133 antibody [EPR20980-45] (ab226355)

All lanes : Anti-CD133 antibody [EPR20980-45] (ab226355) at 1/1000 dilution

Lane 1 : Human pancreas lysate
Lane 2 : NT2/D1 (human malignant pluripotent embryonic carcinoma epithelial cell-like), whole cell lysate
Lane 3 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Developed using the ECL technique.

Predicted band size: 97 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?

Blocking: 5% NFDM/TBST.

Exposure times:

Lane 1: 3 minutes

Lanes 2-3: 10 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR20980-45] (ab226355)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling CD133 with ab226355 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP), ready to use. Apical/endoluminal staining on ducts of human pancreas, and negative on Langerhans cells of the islets (PMID: 18261235) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP), ready to use.
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR20980-45] (ab226355)

Immunohistochemical analysis of paraffin-embedded human pancreatic cancer tissue labeling CD133 with ab226355 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP), ready to use. Apical/endoluminal staining on ducts of human pancreatic cancer (PMID: 18261235) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP), ready to use.
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Anti-CD133 antibody [EPR20980-45] (ab226355)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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