CD45RA antibody (APC)

Catalog No. ABIN1112158

Product Details anti-CD45RA Antibody (APC)

Target: CD45RA

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This CD45RA antibody is conjugated to APC

Application: Flow Cytometry (FACS), Immunofluorescence (IF)

Characteristics: Mouse monoclonal Anti-Human CD45RA APC, is recommended for use in flow cytometry. This product allows simultaneous detection and enumeration of the helper/inducer T-cell subset and CD45RA+ cells.

Clone: HI100

Isotype: IgG1

Application Details

Application Notes: It is recommended for use in flow cytometry. This reagent is effective for direct immunofluorescence staining of human tissue for flow cytometric analysis using 20 µl/10^6 cells.

Sample Preparation:

1. Transfer 100 µl of anticoagulated (EDTA) blood to a 12 x 75 mm polystyrene test tube (10^6 cells). 2. Add 20 µl of CD45RA APC and mix gently with a vortex mixer. The 20 µl is a guideline only, the optimal volume should be determined by the individual laboratory. 3. The recommended negative control is a non-reactive APC-conjugated antibody of the same isotype 4. Incubate in the dark at room temperature at 4°C for 15 minutes or at room temperature (20-25 °C) for 15 minutes. 5. Add 1,5 ml of Lysing Solution to each sample and mix gently with a vortex mixer. Incubate for 10 minutes at room temperature in the dark. 6. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 8. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 9. Resuspend pellet in an appropriate fluid for flow cytometry, e.g. 0.5 ml PBS. The PBS should contain 1% paraformaldehyde (fixative) if samples are not analysed the same day. 10. Analyse on a flow cytometer or store at 2-8 °C in the dark until analysis. Samples can be run up to 24 hours after lysis. Normal Peripheral Blood (SPN) from a Human Donor

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: The conjugate is provided in liquid form in buffer containing and Antibody Stabilizer solution, PBS 20 mM, pH 7,4.

Preservative: Sodium azide

Precaution of Use: 1. The device is not intended for clinical use including diagnosis, prognosis, and monitoring of a disease state, and it must not be used in conjunction with patient records or treatment. 2. This product contains Sodium azide (NaN3), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, Sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 3. As with any product derived from biological sources, proper handling procedures should be used. 7. Add 2 mL 0.01 mol/l PBS (It betters that it containing 2% bovine serum albumin) and resuspend the cells by using a vortex mixer. 0

Storage: 4 °C

References for anti-CD45RA antibody (APC) (ABIN1112158)

Marin, Artal, Martin, Carrizo, Andres, Martin-Burriel, Bolea, Sanz, Varona, Godino, Gallego, Garcia-Erce, Villar, Gil, Forner, Cubero, Ros: "Epigenetics modifications and Subclinical Atherosclerosis in Obstructive Sleep Apnea: The EPIOSA study." in: BMC pulmonary medicine, Vol. 14, pp. 114, (2015) (PubMed).

Target Details for CD45RA

Target: CD45RA

Alternative Name: CD45RA (CD45RA Products)

Synonyms: B220 antibody, CD45 antibody, CD45R antibody, GP180 antibody, L-CA antibody, LCA antibody, LY5 antibody, T200 antibody, Lca antibody, RT7 antibody, protein tyrosine phosphatase, receptor type C antibody, protein tyrosine phosphatase, receptor type, C antibody, PTPRC antibody, Ptprc antibody

Background: CD45 has been identified as a transmembrane glycoprotein, broadly expressed among hematopoietic cells. Multiple isoforms of CD45 are distributed throughout the immune system according to cell type. These isoforms arise because of alternative splicing of exons 4, 5, and 6. The corresponding protein domains are characterized by the binding of monoclonal antibodies specific for CD45RA (exon 4), CD45RB (exon 5), CD45RC (exon 6), and CD45RO (exons 4 to 6 spliced out). The variation in these isoforms is localized to the extracellular domain of CD45, while the intracellular domain is conserved. CD45 functions as a phosphotyrosine phosphatase, a vital component for efficient tyrosine phosphorylation induction by the TCR/CD3 complex. The tyrosine phosphatase activity of CD45 is contained within the conserved intracellular domain. Src and Syk family protein tyrosine kinases are utilized by the TCR/CD3 complex to initiate signaling cascades. Several members of these two families, including Lck, Fyn and ZAP-70, have been implicated as physiological substrates of CD45.