Anti-CD45 Antibody, clone 5L3, ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1180 - recombinant, expressed in HEK 293 cells
Synonym: Anti-B220; Anti-CD45; Anti-CD45R; Anti-GP180; Anti-L-CA; Anti-LCA; Anti-LY5; Anti-T200
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 1:100 dilution of Cat. No. ZRB1180, Anti-CD45, clone 5L3 ZooMAb®, Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in both germinal center & non-germinal center cells of human tonsil tissue.
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 1:100 dilution of Cat. No. ZRB1180, Anti-CD45, clone 5L3 ZooMAb®, Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in both germinal center & non-germinal center cells of human tonsil tissue.
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 1:100 dilution of Cat. No. ZRB1180, Anti-CD45, clone 5L3 ZooMAb®, Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in both germinal center & non-germinal center cells of human tonsil tissue.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and mouse spleen (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1180, Anti-CD45, clone 5L3 ZooMAb®, Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in both white & red pulp of mouse spleen tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and mouse spleen (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1180, Anti-CD45, clone 5L3 ZooMAb®, Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in both white & red pulp of mouse spleen tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and mouse spleen (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1180, Anti-CD45, clone 5L3 ZooMAb®, Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in both white & red pulp of mouse spleen tissue sections.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1180, Anti-CD45, clone 5L3 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 cell membrane.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1180, Anti-CD45, clone 5L3 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 cell membrane.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1180, Anti-CD45, clone 5L3 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 cell membrane.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Lysates from Jurkat cells (Lane 1) and rat spleen (Lane 2) were probed with Cat. No. ZRB1180, Anti-CD45, clone 5L3 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 CD45 (~200 kDa).
Western Blotting Enhanced Validation-Recombinant Antibody Technology Human tonsil lysate was probed with Cat. No. ZRB1180, Anti-CD45, clone 5L3 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 CD45 (~200 kDa).
Flow Cytometry Enhanced Validation-Recombinant Antibody Technology Staining of one million jurkat cells was performed using 1μg of Cat. No. ZRB1180, Anti-CD45, clone 5L3 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-Rabbit IgG 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 | 5L3, 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 61.01 kDa |
| observed mol wt ~200 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | rat, human, mouse |
| storage temp. | 2-8°C |
| technique(s) | flow cytometry: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable (peptide) | |
| UniProt accession no. | P08575  |
| Application: | Anti-CD45, clone 5L3 ZooMAb , Cat. No. ZRB1180, is a highly specific recombinant rabbit monoclonal antibody that detects CD45 and has been tested for use in Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected CD45 in rat spleen and human tonsil tissue lysate. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected CD45 in human tonsil and mouse spleen tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected CD45 in Jurkat cells. Note: Actual optimal working dilutions must be determined by the 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: | KLH-conjugated linear peptide corresponding to 16 amino acids from the C-terminal, cytoplasmic domain of human CD45. |
| 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 is 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 5L3 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects CD45. It targets an epitope within 16 amino acids from the C-terminal half, cytoplasmic domain. |
| 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: | Receptor-type tyrosine-protein phosphatase C (UniProt: P08575; also known as EC:3.1.3.48, Leukocyte common antigen, L-CA, T200, CD45) is encoded by the PTPRC (also known as CD45) gene (Gene ID: 5788) in human. CD45 is a single-pass type I membrane protein with tyrosine phosphatase activity. CD45 has been shown to be essential for antigen-induced signaling in T-cells and B-cells. It acts as a positive regulator of T-cell coactivation upon binding to DPP4. It is synthesized with a signal peptide (aa 1-25), which is subsequently cleaved off to generate the mature form that has a extracellular domain (aa 26-577), a short transmembrane domain (aa 578-598), and a cytoplasmic domain (aa 599-1306). CD45 contains two protein tyrosine phosphatase (PTPase) domains (aa 653-912 and 944-1228) and the first PTPase domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. It also contains 2 fibronectin type-III domains (aa 391-483 and 484-576). Upon T-cell activation, it recruits and dephosphorylates SKAP1 and FYN. It also dephosphorylates LYN, and thereby modulates its activity. CD45 is expressed at high levels on all hematopoietic cells, however, higher density expression occurs on lymphocytes where about 10% of the surface area may be CD45. Eight isoforms of CD45 have been described that are produced by alternative splicing. However, only isoforms 1,2,3,5, and 7 are detected in thymocytes. Mutations in PTPRC gene have been associated with Multiple Sclerosis. 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 |