WB result of CD58 Rabbit mAb
Primary antibody: CD58 Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: Raji whole cell lysate 20 µg
Lane 3: Jurkat whole cell lysate 20 µg
Lane 4: K562 whole cell lysate 20 µg
Lane 5: SH-SY5Y whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 28 kDa
Observed MW: 70 kDa
CD58 Recombinant Rabbit mAb (S-R295)
CD58 Recombinant Rabbit mAb (S-R295)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | CD58 |
| Synonyms | Lymphocyte function-associated antigen 3, Ag3, Surface glycoprotein LFA-3, LFA3 |
| Location | Cell membrane |
| Accession | P19256 |
| Clone Number | S-R295 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC, ICFCM |
| Reactivity | Hu |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05%BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | |
| ICC | 1:500 | |
| ICFCM | 1:500 |
Background
CD58, or lymphocyte function-associated antigen 3 (LFA-3), is a cell adhesion molecule expressed on Antigen Presenting Cells (APCs), particularly macrophages, and other tissue cells. CD58 binds to CD2 (LFA-2) on T cells and is important in strengthening the adhesion and recognition between the T cells and Professional Antigen Presenting Cells, facilitating signal transduction necessary for an immune response. CD58 also plays a role in the regulation of colorectal tumor-initiating cells (CT-ICs). Thus, cells that express CD58 have become a cell of interest in tumorigenesis. Mutations of CD58 have been linked to immune evasion observed in some lymphomas and studies are underway to analyze how its involvement directly affects classical Hodgkin lymphoma (cHL).
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Western Blot
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte) labelling CD58 antibody at 1/500 dilution (0.1 μg) / (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Immunocytochemistry
ICC shows positive staining in Jurkat cells. Anti-CD58 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
