Immunohistochemical analysis of paraffin-embedded Human Hodgkin's lymphoma tissue labeling CD33 with ab269456 at 1/200 dilution (2.96 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on human Hodgkin's lymphoma.

The section was incubated with ab269456 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

CD33 was immunoprecipitated from 0.35 mg THP-1 (Human monocytic leukemia monocyte) whole cell lysate with ab269456 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab269456 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: THP-1 whole cell lysate 10ug.

Lane 2: ab269456 IP in THP-1 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab269456 in THP-1 whole cell lysate.

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

Exposure time: 23 seconds.

 

Immunofluorescent analysis of 100% methanol-fixed THP-1 (human monocytic leukemia monocyte) and Jurkat (human T cell leukemia T lymphocyte) cells labeling CD33 with ab26946 at 1/50 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution (Green). Confocal image showing membranous staining in THP-1 cells. The nuclear counter stain is DAPI (Blue). Tubilin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red).

Control: Used PBS instead of primary antibody, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

Negative control: Jurkat (PMID: 30519686).

 

All lanes : Anti-CD33 antibody [EPR23051-101] (ab269456) at 1/1000 dilution

Lane 1 : U937 (human histiocytic lymphoma monocyte), whole cell lysate
Lane 2 : THP-1 (human monocytic leukemia monocyte), whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 40 kDa
Observed band size: 67/75 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 70 seconds.

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

Negative Control:Jurkat (PMID:30519686).

All lanes : Anti-CD33 antibody [EPR23051-101] (ab269456) at 1/1000 dilution

Lane 1 : Human liver tissue lysate
Lane 2 : Human skeletal muscle tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

Predicted band size: 40 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 70 seconds.

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

Negative control:Human skeletal muscle.

Immunohistochemical analysis of paraffin-embedded Human clear cell renal cell carcinoma tissue labeling CD33 with ab269456 at 1/200 dilution (2.96 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on infiltrated myeloid cells of human clear cell renal cell carcinoma The section was incubated with ab269456 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling CD33 with ab269456 at 1/200 dilution (2.96 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on mainly Kuffer cells of human liver (PMID:25721896). The section was incubated with ab269456 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling CD33 with ab269456 at 1/200 dilution (2.96 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Negative control：Human skeletal muscle.

The section was incubated with ab269456 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.