Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (AB206127)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic adenocarcinoma tissue labelling CD166 with unpurified ab109215 at 1/100.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (AB206127)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD166 with unpurified ab109215 at 1/100.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt

Lab

Flow Cytometry - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (AB206127)

Flow Cytometry analysis of HuT-78 cells labelling CD166 with purified ab109215 at 1/90 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (AB206127)

Immunocytochemistry/Immunofluorescence analysis of THP-1 (human monocytic leukemia cell line) cells labelling CD166 (green) with purified ab109215 at 1/250. Cells were fixed with 100% methanol. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as a nuclear counterstain.

Secondary Only Control : PBS was used instead of the primary antibody as the negative control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (AB206127)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD166 with purified ab109215 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

• IP

Lab

Immunoprecipitation - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (AB206127)

ab109215 (purified) at 1/30 immunoprecipitating CD166 in SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

All lanes:

Immunoprecipitation - Anti-CD166 antibody [EPR2759(2)] (<a href='/en-us/products/primary-antibodies/cd166-antibody-epr27592-ab109215'>ab109215</a>)

Predicted band size: 65 kDa

Observed band size: 100-105 kDa

false

• WB

Lab

Western blot - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (AB206127)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : CD166 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : SH-SY5Y whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab109215 observed at 100 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab109215 was shown to specifically react with CD166 in wild-type HAP1 cells as signal was lost in CD166 knockout cells. Wild-type and CD166 knockout samples were subjected to SDS-PAGE. ab109215 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD166 antibody [EPR2759(2)] (<a href='/en-us/products/primary-antibodies/cd166-antibody-epr27592-ab109215'>ab109215</a>)

Predicted band size: 65 kDa

false

• WB

Unknown

Western blot - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (AB206127)

Blocking buffer and concentration : 5% NFDM/TBST

Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)

All lanes:

SH-SY5Y (human neuroblastoma) whole cell lysate at 10 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>)

Predicted band size: 65 kDa

Observed band size: 100-105 kDa

false

Exposure time: 1min