Anti-CD133 antibody [EPR20980-45]

• IHC-P

Supplier Data

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).

• IHC-P

Supplier Data

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).

• WB

Supplier Data

Western blot - Anti-CD133 antibody [EPR20980-45] (AB226355)

Blocking : 5% NFDM/TBST.

Exposure times :

Lane 1 : 3 minutes

Lanes 2-3 : 10 seconds

All lanes:

Western blot - Anti-CD133 antibody [EPR20980-45] (ab226355) at 1/1000 dilution

Lane 1:

Human pancreas lysate at 10 µg

Lane 2:

NT2/D1 (human malignant pluripotent embryonic carcinoma epithelial cell-like), whole cell lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Secondary

Lanes 1 - 2:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lane 3:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 97 kDa

Observed band size: 120 kDa

true

• WB

Lab

Western blot - Anti-CD133 antibody [EPR20980-45] (AB226355)

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.

All lanes:

Western blot - 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

Predicted band size: 97 kDa

Observed band size: 110 kDa

false

Exposure time: 20min

• WB

Supplier Data

Western blot - Anti-CD133 antibody [EPR20980-45] (AB226355)

Blocking : 5% NFDM/TBST.

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

Negative control : HEK-293T (PMID 20167130).

All lanes:

Western blot - Anti-CD133 antibody [EPR20980-45] (ab226355) at 1/1000 dilution

Lane 1:

Caco-2 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

HT-29 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

NCCIT (human pluripotent embryonic carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 97 kDa

Observed band size: 120 kDa

true

Exposure time: 15s