Anti-CD54 Antibody, clone 2C15 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1296 - recombinant, expressed in HEK 293 cells
Synonym: Intercellular adhesion molecule 1;ICAM-1;Major group rhinovirus receptor
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human lung (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1296, Anti-CD54, clone 2C15 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining in alveoli of human lung tissue sections was observed.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human lung (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1296, Anti-CD54, clone 2C15 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining in alveoli of human lung tissue sections was observed.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human lung (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1296, Anti-CD54, clone 2C15 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining in alveoli of human lung tissue sections was observed.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from Human Liver tissue (Lane 1), Ramos cells (Lane 2) and MCF-7 cells (Lane 3) were probed with Cat. No. ZRB1296, Anti-CD54, clone 2C15 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 CD54 (~100 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or CD54 (ICAM-1) peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1296, Anti-CD54, clone 2C15 ZooMAb® Rabbit Monoclonal. The binding of this antibody to ICAM-1/CD54 peptide displayed a KD of 2.0 x 10-8, whereas binding to non-specific control peptide displayed a KD of 1.5 x 10-4.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million Human peripheral blood mononuclear cells (PBMC) was performed using 1 μg of Cat. No. ZRB1296, Anti-CD54, clone 2C15 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 |
| clone | 2C15, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Extracellular domain |
| 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 57.82 kDa |
| observed mol wt ~100 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_000192  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | monkey |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| flow cytometry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | P05362 |
| Application: | Quality Control Testing Evaluated by Western Blotting in Ramos cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected CD54 in Ramos cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected CD54 in lysates from human liver tissue and MCF-7 cells. Flow Cytometry Analysis: 1 μg from a representative lot detected CD54 in one million human peripheral blood mononuclear cells. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected CD54 in human lung tissue sections. Affinity Binding Assay: A representative lot of this antibody bound ICAM-1/CD54 with a KD of 2.0 x 10-8 in an affinity binding assay. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| 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. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 18 amino acids from the N-terminal, extracellular domain of human CD54 (ICAM-1). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 2C15 is a Rabbit recombinant monoclonal antibody that specifically detects CD54. It targets an epitope within 18 amino acids from the N-terminal, extracellular 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: | Intercellular adhesion molecule 1 (UniProt: P05362; also known as ICAM-1, Major group rhinovirus receptor, CD54) is encoded by the ICAM1 gene (Gene ID: 3383) in human. CD54 is a member of the immunoglobulin supergene family with five extracellular Ig like domains that are involved in cell-cell and cell-matrix adhesion. It is a single-pass type I membrane glycoprotein that is synthesized with a signal peptide (aa 1-27), which is subsequently cleaved off to generate the mature form that contains an extracellular domain (aa 28-481), a transmembrane domain (aa 481-503), and a cytoplasmic domain (aa 504-532). It is reported to be constitutively expressed in several hematopoietic and non-hematopoietic cells, including fibroblasts, endothelial cell, leukocytes, and epithelial cells. Its expression can be upregulated by several pro-inflammatory cytokines. Stronger expression of CD54 has been observed in neoplastic cells from stem cell derived neoplasms. The endothelial expression of CD54 is showed to be higher in atherosclerotic and transplant-associated atherosclerotic tissue. Its adhesive actions are mediated by binding to CD11a/CD18 (LFA-1) and CD11b/CD18 (Mac-1). Its adhesive interactions are shown to be essential for the transendothelial migration of leukocytes out of the blood vessels and into tissues. It also plays a role in the activation of T cells where it serves as a co-activation signal. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Lawson, C., and Wolf, S (2009). Pharmacol Rep. 61(1); 22-32; Roebuck, KA., and Finnegan, A. (1999). J. Leukoc. Biol. 66(6); 876-888; Maio, M., and Del Vecchio, L (1992). Leuk. Lymphoma. 8(2-2); 23-33). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |