Anti-CD31 antibody [RM1006]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [RM1006] (AB281583)

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-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [RM1006] (AB281583)

Immunohistochemical analysis of formalin fixed paraffin embedded Human placenta labelling CD31 with ab281583 at a concentration of 0.5µg/ml.

The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5. ab281583 anti-CD31 was incubated at 37°C for 16min.

Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

AbReview80744****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [RM1006] (AB281583)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of paraformaldehyde fixed human tonsil permeabilized with 0.3% Triton X-100, staining with ab281583 at 10µg/ml. Secondary antibody used was ab150073 at 1/500 dilution. Blocking was done using Donkey Serum 10% + 3% BSA for 24 hours at 4°C. The sample was incubated with the primary antibody for 1 hour at 20°C. Validated on GE Cell DIVE. Antigen retrieval method was heat mediated Citrate pH 6 & TRIS pH 9

This image is courtesy of an Abreview submitted by Dylan Windell

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [RM1006] (AB281583)

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.

• IHC-P

AbReview81814****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [RM1006] (AB281583)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skin tissue staining CD13 with ab281583 at 1/1000 dilution and secondary Alexa Fluor® 488 Goat anti-rabbit IgG (H+L) at 1/300 dilution. The tissue was incubated with the primary antibody for 12 hour at 4 degrees. Blocking was done with 5% BSA for 30 minutes at 21 degrees. Heat mediated antigen retrieval was with Tris/EDTA buffer pH 9.0

This image is courtesy of an anonymous Abreview

• IP

Supplier Data

Immunoprecipitation - Anti-CD31 antibody [RM1006] (AB281583)

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.

All lanes:

Immunoprecipitation - Anti-CD31 antibody [RM1006] (ab281583)

Predicted band size: 82 kDa

Observed band size: 125 kDa

false

• IHC-P

AbReview81813****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [RM1006] (AB281583)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of paraformaldehyde-fixed mouse skin tissue staining with ab281583 at 1/4000 dilution. Secondary antibody was HRP-Goat Anti-Rabbit IgG (H+L) at 1/500 dilution. Samples were incubated with the primary antibody for 12 hours at 4°C. Blocking was done using 5% BSA for 30 minutes at 21°C. Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0.

This image is courtesy of an anonymous Abreview

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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.

• IP

Supplier Data

Immunoprecipitation - Anti-CD31 antibody [RM1006] (AB281583)

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.

All lanes:

Immunoprecipitation - Anti-CD31 antibody [RM1006] (ab281583)

Predicted band size: 82 kDa

Observed band size: 125 kDa

false

• WB

Lab

Western blot - Anti-CD31 antibody [RM1006] (AB281583)

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

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

Exposure time : 37 seconds

All lanes:

Western blot - Anti-CD31 antibody [RM1006] (ab281583) at 1/1000 dilution

Lane 1:

HUVEC (Human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 2:

bEnd.3 (Mouse brain endothelioma ) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

Rat lung 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 1/100000 dilution

Predicted band size: 82 kDa

Observed band size: 125 kDa

false