Anti-CD31 antibody [EPR3094]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (AB76533)

ab76533 staining CD31 in human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (AB76533)

Immunohistochemical analysis of formalin fixed paraffin embedded human kidney labelling CD31 with ab76533 at a concentration of 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5 for 32mins.

ab76533 anti-CD31 [EPR3094] was incubated for 16mins at 37°C. Sections were counterstained 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

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (AB76533)

Immunohistochemical analysis of endothelial colony forming progenitor cell plugs, staining CD31 (green) with ab76533.

Following antigen retrieval and blocking, sections were incubated with primary antibody (1/1000) overnight at 4°C. A Cy5®-conjugated anti-rabbit IgG (2 μg/ml) was used as the secondary antibody.

Image from Hofmann NA et al. PLoS One. 2012;7(9):e44468. doi: 10.1371/journal.pone.0044468. Epub 2012 Sep 7. Fig 3.; doi:10.1371/journal.pone.0044468; September 7 2012 PLoS ONE 7(9): e44468.

• IHC-P

AbReview23029****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (AB76533)

ab76533 staining CD31 in human muscle tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then blocked with 3% serum for 30 minutes at 20°C followed by incubation with the primary antibody at a 1/200 dilution for 12 hours at20°C. A Cy3®-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/200 dilution.

This image is courtesy of an anonymous Abreview.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (AB76533)

Tissue Microarrays stained for "Anti-CD31 antibody [EPR3094]" using "ab76533"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab76533 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CD31 antibody [EPR3094] (AB76533)

ab76533 staining CD31 in THP-1 (Human monocytic leukemia monocyte) cell line. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab76533 at 1/250 (4.9 μ μg/mL) overnight at +4°C. ab195889Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used as a counterstain at 1/200 dilution. DAPI (blue) was used as nuclear counterstain. Confocal image showing membranous staining in THP-1 cells.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD31 antibody [EPR3094] (AB76533)

This image was generated using ab227688, the same clone, but with a different buffer formulation. Panel A : merged staining of anti-CD31 (gray; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD19 (red; Opal™570) on Formalin/PFA-fixed paraffin-embedded sections of human tonsil. Secondary antibody was Opal Polymer HRP Ms + Rb, nuclear counterstain was DAPI. Panel B : anti-CD3 epsilon stained on T cells with ab16669 at 1/500 dilution Panel C : anti-CD19 stained on B cells with ab237772 at 1/5000 dilution Panel D : anti-CD31 stained on endothelial cells and immune cell subsets with ab207090 at 1/500 dilution The section was incubated in three rounds of staining : in the order of ab207090 and ab16669 for 30 mins, then ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

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Western blot - Anti-CD31 antibody [EPR3094] (AB76533)

All lanes:

Western blot - Anti-CD31 antibody [EPR3094] (ab76533) at 1/20000 dilution

Lane 1:

THP-1 cell lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 82 kDa

Observed band size: 125 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (AB76533)

Immunohistochemical analysis of paraffin embedded human kidney tissue using ab76533 at a 1/250 dilution. Note positive staining of endothelial cells.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (AB76533)

Immunohistochemical analysis of paraffin embedded human muscle tissue using ab76533 at a 1/250 dilution. Note positive staining of endothelial cells.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-CD31 antibody [EPR3094] (AB76533)

Blocking and diluting buffer : 5% NFDM /TBST

All lanes:

Western blot - Anti-CD31 antibody [EPR3094] (ab76533) at 1/10000 dilution

All lanes:

THP-1 (Human monocytic leukemia cell line) 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 1/20000 dilution

Predicted band size: 82 kDa

Observed band size: 125 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-CD31 antibody [EPR3094] (AB76533)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.