Anti-CD133 antibody [EPR24348-38]

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CD133 antibody [EPR24348-38] (AB271092)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse E14.5 embryonic kidney tissue labeling CD133 with ab271092 at 1/100 (4.59 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on the apical membrane of mouse E14.5 embryonic kidney is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CD133 antibody [EPR24348-38] (AB271092)

Flow Cytometry analysis of Mouse bone marrow cells labelling CD133 with ab271092 at 1/50 (right). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) was used as the isotype control (left). Cells were co-stained with anti-CD117 conjugated to BV421. Gated on viable cells.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CD133 antibody [EPR24348-38] (AB271092)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse E14.5 embryonic lung tissue labeling CD133 with ab271092 at 1/100 (4.59 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on the apical membrane of lung buds is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR24348-38] (AB271092)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labelling CD133 with ab271092 at 1/2000 (0.23 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse kidney. The section was incubated with ab271092 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CD133 antibody [EPR24348-38] (AB271092)

Flow Cytometry analysis of Mouse bone marrow cells labelling CD133 with ab271092 at 1/50 (right). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat Anti-Rabbit IgG (Alexa Fluor® 647) (ab150083) at 1/2000 was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) was used as the isotype control (left). Cells were stained with rabbit IgG or ab271092. Then stained with Lineage cocktail conjugated to FITC, anti-Sca-1 conjugated to PE/Cy7, anti-CD117 conjugated to APC/Cy7 and anti-CD16/32 conjugated to BV510.
Gated on viable MP cells. Positive staining on CD16/32+ MP cells is consistent with literature. (PMID : 23509298).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CD133 antibody [EPR24348-38] (AB271092)

Flow Cytometry analysis of Mouse bone marrow cells labelling CD133 with ab271092 at 1/50 (right). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) was used as the isotype control (left). Gated on viable cells.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR24348-38] (AB271092)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling CD133 with ab271092 at 1/2000 (0.23 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the biliary ducts of mouse liver. The section was incubated with ab271092 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR24348-38] (AB271092)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labelling CD133 with ab271092 at 1/2000 (0.23 ug/ml) followed by a ready to use LeicaDS9800(Bond™ Polymer Refine Detection). Positive staining on mouse pancreatic ducts. The section was incubated with ab271092 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [EPR24348-38] (AB271092)

Immunohistochemical analysis of paraffin-embedded Mouse retina tissue labelling CD133 with ab271092 at 1/2000 (0.23 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the cone and rod cells of mouse retina. The section was incubated with ab271092 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CD133 antibody [EPR24348-38] (AB271092)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse E14.5 embryonic cortex tissue labeling CD133 with ab271092 at 1/100 (4.59 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on the apical membrane of mouse E14.5 embryonic cortex is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IP

Supplier Data

Immunoprecipitation - Anti-CD133 antibody [EPR24348-38] (AB271092)

CD133 was immunoprecipitated from 0.35 mg Mouse E14.5 brain tissue lysate 10 ug with ab271092 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271092 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse E14.5 brain tissue lysate 10 ug

Lane 2 : ab271092 IP in Mouse E14.5 brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab271092 in Mouse E14.5 brain tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds

All lanes:

Immunoprecipitation - Anti-CD133 antibody [EPR24348-38] (ab271092)

Predicted band size: 97 kDa

Observed band size: 120 kDa

false

• WB

Lab

Western blot - Anti-CD133 antibody [EPR24348-38] (AB271092)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : mouse heart (PMID : 17575139).

Exposure time : Lanes 1-2 : 10 seconds; Lane 3 : 70 seconds.

All lanes:

Western blot - Anti-CD133 antibody [EPR24348-38] (ab271092) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse E14.5 brain tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/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

false

• WB

Lab

Western blot - Anti-CD133 antibody [EPR24348-38] (AB271092)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-CD133 antibody [EPR24348-38] (ab271092) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate

Lane 2:

Mouse kidney tissue lysate treated with PNGase F

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/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,97 kDa

false

Exposure time: 10s