Recombinant Anti-CD31 antibody [RM1006] (ab281583)

IHC image of ab281583 staining CD31 in rat lung formalin fixed paraffin embedded tissue sections, performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The section was then incubated with ab281583 at 1:4000 dilution (0.116 μg/ml) for 30 mins at room temperature and detected using an HRP conjugated compact polymer system (Bond™ Polymer Refine Detection). DAB was used as the chromogen. Positive staining was obtained on blood vessels in rat lung. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

IHC image of ab281583 staining CD31 in mouse lung formalin fixed paraffin embedded tissue sections, performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The section was then incubated with ab281583 at 1:4000 dilution (0.116 μg/ml) for 30 mins at room temperature and detected using an HRP conjugated compact polymer system (Bond™ Polymer Refine Detection). DAB was used as the chromogen. Positive staining was obtained on blood vessels in mouse lung. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

IHC image of ab281583 staining CD31 in human placenta formalin fixed paraffin embedded tissue sections, performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The section was then incubated with ab281583 at 1:4000 dilution (0.116 μg/ml) for 30 mins at room temperature and detected using an HRP conjugated compact polymer system (Bond™ Polymer Refine Detection). DAB was used as the chromogen. Positive staining was obtained on blood vessels in human placenta. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung tissue labeling CD31 with 281583 at 1/50 (9.24 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse lung 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).

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat lung tissue labeling CD31 with 281583 at 1/50 (9.24 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on rat lung 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).

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

Negative control: NIH/3T3 (PMID: 1429859).

Exposure time: 37 seconds

CD31 was immunoprecipitated from 0.35 mg THP-1 (Human monocytic leukemia monocyte) whole cell lysate 10 ug with 281583 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using 281583 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: THP-1 (Human monocytic leukemia monocyte) whole cell lysate 10 ug

Lane 2: ab281583 IP in THP-1 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab281583 in THP-1 whole cell lysate

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

Exposure time: 6 seconds.

CD31 was immunoprecipitated from 0.35 mg bEnd.3 (Mouse brain endothelioma ) whole cell lysate 10 ug with 281583 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using 281583 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: bEnd.3 (Mouse brain endothelioma ) whole cell lysate 10 ug

Lane 2: ab281583 IP in bEnd.3 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab281583 in bEnd.3 whole cell lysate

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

Exposure time: 6 seconds.