Anti-CD31 Antibody, clone 4E20 , ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1216 - recombinant, expressed in HEK 293 cells
Synonym: Platelet endothelial cell adhesion
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and human tonsil (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10,000 dilution of Cat. No. ZRB1216, Anti-CD31, clone 4E20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in endothelium of human tonsil sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and human tonsil (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10,000 dilution of Cat. No. ZRB1216, Anti-CD31, clone 4E20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in endothelium of human tonsil sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and human tonsil (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10,000 dilution of Cat. No. ZRB1216, Anti-CD31, clone 4E20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in endothelium of human tonsil sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of THP-1 cell line were performed using a 1:100 dilution of Cat. No. ZRB1216, Anti-CD31, clone 4E20 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasmic domain of plasma membrane.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of THP-1 cell line were performed using a 1:100 dilution of Cat. No. ZRB1216, Anti-CD31, clone 4E20 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasmic domain of plasma membrane.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of THP-1 cell line were performed using a 1:100 dilution of Cat. No. ZRB1216, Anti-CD31, clone 4E20 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasmic domain of plasma membrane.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Human spleen (Lane 1) and HL-60 cell (Lane 2) lysates were probed with Cat. No. ZRB1216, Anti-CD31, clone 4E20 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates CD31 (~130 kDa).
Western Blotting Enhanced Validation - Recombinant Antibody Technology THP-1 (Lane 1) and Jurkat (Lane 2) cell lysates were probed with Cat. No. ZRB1216, Anti-CD31, clone 4E20 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates CD31 (~130 kDa).
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million THP-1 cell line was performed using 0.2 μg/mL of Cat. No. ZRB1216, Anti-CD31, clone 4E20 ZooMAb® Rabbit Monoclonal (yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (grey/white histogram), followed by a PE-conjugated Donkey Anti-Rabbit secondary antibody.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit (recombinant) |
| clone | 4E20, recombinant monoclonal |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 82.52 kDa |
| observed mol wt 130 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| storage temp. | 2-8°C |
| technique(s) | flow cytometry: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry: suitable | |
| western blot: suitable | |
| UniProt accession no. | P16284  |
| Application: | Anti-CD31, clone 4E20 ZooMAb, Cat. No. ZRB1216, is a highly specific rabbit recombinant monoclonal antibody that targets Platelet endothelial cell adhesion molecule (CD31 / PECAM-1) and has been tested for use in Flow Cytometry, Immunocytochemistry, Immunohistochemistry, and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected CD31 in THP1, HL60, and Jurkat cell lysates. Immunohistochemistry (Paraffin) Analysis: A 1:10,000 dilution from a representative lot detected CD31 in human tonsil tissue sections. Flow Cytometry Analysis: 0.2 μg/mL from a representative lot detected CD31 in THP-1 cells. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected CD31 in THP-1 cell line. Note: Actual optimal working dilutions must be determined by end user as specimens and experimental conditions may vary. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | GST/His-tagged recombinant fragment corresponding to 85 amino acids from the C-terminal, cytoplasmic domain of human Platelet endothelial cell adhesion molecule (CD31; PECAM-1). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 30 μg/mL after reconstitution at 25 μL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 4E20 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects human Platelet endothelial cell adhesion molecule (PECAM-1). It targets an epitope within the cytoplasmic domain from the C-terminal region. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Platelet endothelial cell adhesion molecule (UniProt: P16284; also known as PECAM-1, EndoCAM. GPIIA′, PECA1, CD31) is encoded by the PECAM1 gene (Gene ID: 5175) in human. PECAM-1 is a single-pass type I membrane protein that is synthesized with a signal peptide (aa 1-27) that is subsequently cleaved to generate the mature protein with a large extracellular region (aa 28-601) that contains six Ig-like C2-type domains, followed by a transmembrane segment (aa 602-620) and a cytoplasmic tail (aa. 621-738 of isoform Long) that plays an essential role in PECAM-1-mediated cellular signaling. It contains six Ig-like C2-type domains and Ig-like C2-type domains 2 and 3 contribute to formation of the complex with BDKRB2 and in regulation of its activity. PECAM-1 is essential for leukocyte transendothelial migration (TEM) under most inflammatory conditions. It is expressed on platelets and leukocytes and is primarily concentrated at the borders between endothelial cells. Six isoforms of PECAM-1 have been described that are produced by alternative splicing. Isoform Long predominates in all tissues examined, whereas Delta 12 isoform is detected only in trachea and isoform Delta14-15 are only detected in lung tissue. Isoform Delta14 is detected in all tissues examined with the strongest expression in heart. Isoform Delta15 is expressed in brain, testis, ovary, cell surface of platelets, human umbilical vein endothelial cells (HUVECs), Jurkat T-cell leukemia, human erythroleukemia and U-937 histiocytic lymphoma cell lines. Tyrosine 690 residue plays a critical role in TEM and is required for efficient trafficking of PECAM-1 to and from the lateral border recycling compartment (LBRC) and is also required for the LBRC membrane to be targeted around migrating leukocytes. PECAM-1 also prevents phagocyte ingestion of closely apposed viable cells by transmitting ′detachment′ signals, and changes function on apoptosis, promoting tethering of dying cells to phagocytes. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |