Anti-CD133 antibody [EPR20980-104]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR20980-104] (AB216323)

Immunohistochemical analysis of paraffin-embedded Human pancreatic carcinoma tissue labelling CD133 with ab216323 at 1/1000 dilution (1.386 μg/ml) followed by a Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880) at Ready to use dilution. Positive staining on human pancreatic carcinoma. The section was incubated with ab216323 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880) at Ready to use dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR20980-104] (AB216323)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CD133 with ab216323 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP), ready to use. Membranous staining on parietal layer of Bowman's capsule of human kidney (PMID : 19092120) 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 [EPR20980-104] (AB216323)

Immunohistochemical analysis of paraffin-embedded human mammary gland tissue labeling CD133 with ab216323 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP), ready to use. Apical/endoluminal staining on ducts of human mammary gland (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 [EPR20980-104] (AB216323)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling CD133 with ab216323 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).

• Flow Cyt

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Flow Cytometry - Anti-CD133 antibody [EPR20980-104] (AB216323)

Flow cytometric analysis of HEK-293T (human embryonic kidney epithelial cell)(Left) / Caco-2 (human colorectal adenocarcinoma epithelial cell)(Right) cell line labeling CD133 with ab216323 at 1/100 (Red) compared with Isotype Control Rabbit monoclonal IgG (ab172730) (BLack) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

Negative control : 293T (PMID 20167130). Total viable cells were gated for the FC image.

• IP

Supplier Data

Immunoprecipitation - Anti-CD133 antibody [EPR20980-104] (AB216323)

CD133 was immunoprecipitated from 0.35 mg Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate with ab216323 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab216323 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution
Lane 1 : Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate 10 μg (Input).
Lane 2 : ab216323 IP in Caco-2 whole cell lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab216323 in Caco-2 whole cell lysate (-).
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-CD133 antibody [EPR20980-104] (ab216323)

Predicted band size: 97 kDa

Observed band size: 120 kDa

false

• WB

Lab

Western blot - Anti-CD133 antibody [EPR20980-104] (AB216323)

ab216323 was shown to react with CD133 in western blot. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab216323 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 [EPR20980-104] (ab216323) at 1/1000 dilution

Lane 1:

Caco-2 cell lysate at 20 µg

Lane 2:

HeLa cell lysate at 20 µg

Predicted band size: 97 kDa

Observed band size: 110 kDa

false

Exposure time: 8min

• WB

Supplier Data

Western blot - Anti-CD133 antibody [EPR20980-104] (AB216323)

Blocking : 5% NFDM/TBST.

Exposure time :

Lane 1 : 30 seconds

Lane 2 : 1 minute

All lanes:

Western blot - Anti-CD133 antibody [EPR20980-104] (ab216323) at 1/1000 dilution

Lane 1:

NCCIT (human pluripotent embryonic carcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

Human fetal kidney at 10 µg

Secondary

Lane 1:

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

Lane 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 1/20000 dilution

Predicted band size: 97 kDa

Observed band size: 120 kDa

true

• WB

Supplier Data

Western blot - Anti-CD133 antibody [EPR20980-104] (AB216323)

Blocking : 5% NFDM/TBST.

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

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

All lanes:

Western blot - Anti-CD133 antibody [EPR20980-104] (ab216323) at 1/1000 dilution

Lane 1:

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

Lane 2:

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: 3min