Anti-CD33 antibody [EPR24370-124] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR24370-124] - BSA and Azide free (AB281568)

This data was developed using ab270942, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining on immune cells of human colon (PMID : 31462392). The section was incubated with ab270942 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-CD33 antibody [EPR24370-124] - BSA and Azide free (AB281568)

This data was developed using ab270942, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining in human skeletal muscle. The section was incubated with ab270942 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.

• WB

Lab

Western blot - Anti-CD33 antibody [EPR24370-124] - BSA and Azide free (AB281568)

This data was developed using ab270942, the same antibody clone in a different buffer formulation. False colour image of Western blot : Anti-CD33 antibody [EPR24370-124] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab270942 was shown to bind specifically to CD33. A band was observed at 60-80 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CD33 knockout cell line ab273831 (knockout cell lysate ab273785). To generate this image, wild-type and CD33 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-CD33 antibody [EPR24370-124] (<a href='/en-us/products/primary-antibodies/cd33-antibody-epr24370-124-ab270942'>ab270942</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 10 µg

Lane 2:

Western blot - Human CD33 knockout THP-1 cell lysate (ab273785)

Lane 2:

CD33 knockout THP-1 cell lysate at 10 µg

Lane 3:

HL-60 cell lysate at 10 µg

Lane 4:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 40 kDa

false

• WB

Lab

Western blot - Anti-CD33 antibody [EPR24370-124] - BSA and Azide free (AB281568)

This data was developed using ab270942, the same antibody clone in a different buffer formulation.

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

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 16380601, 30519686).

Negative Control : Jurkat (PMID : 30519686).

Exposure time : Lanes 1-3 : 136 seconds; Lane 4 : 26 seconds.

All lanes:

Western blot - Anti-CD33 antibody [EPR24370-124] (<a href='/en-us/products/primary-antibodies/cd33-antibody-epr24370-124-ab270942'>ab270942</a>) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 3:

HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg

Lane 4:

TF-1 (human erythroleukemia erythroblast) 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 1/50000 dilution

Predicted band size: 40 kDa

Observed band size: 67-75 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR24370-124] - BSA and Azide free (AB281568)

This data was developed using ab270942, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining in human tonsil. The section was incubated with ab270942 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-CD33 antibody [EPR24370-124] - BSA and Azide free (AB281568)

This data was developed using ab270942, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human Hodgkin's lymphoma tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining in human Hodgkin's lymphoma. The section was incubated with ab270942 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-CD33 antibody [EPR24370-124] - BSA and Azide free (AB281568)

This data was developed using ab270942, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining on Kupffer cells of human liver (PMID : 25721896). The section was incubated with ab270942 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-CD33 antibody [EPR24370-124] - BSA and Azide free (AB281568)

This data was developed using ab270942, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human clear cell renal cell carcinoma tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining on scattered cells of human clear cell renal cell carcinoma. The section was incubated with ab270942 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.