Alexa Fluor® 647 Rat Anti-Mouse CD53
Clone OX-79 (RUO)
- Brand BD Pharmingen™
- Alternative Name Cd53: Ox-44; Tspan25; Tetraspanin-25
- Concentration 0.2 mg/ml
- Isotype Rat AO IgM, κ
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen BAB/14 mouse macrophage cell line RAW 264
- Storage Buffer Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The OX-79 antibody reacts with CD53, a 35-45-kDa member of the Transmembrane 4-pass protein superfamily (TM4SF). As in the human, mouse peripheral leukocytes express CD53 mRNA and protein, and erythrocytes and nonhematopoietic cells do not. However, the distribution of CD53 in the mouse and rat thymus differs from that in the human. In the mouse, most CD4+8- and major subsets of CD4- CD8- (double-negative) and CD4- CD8+ thymocytes express CD53, while most cortical CD4+ CD8+ (double-positive) thymocytes are CD53-low or CD53-negative. CD53 expression can be induced in double-positive thymocytes by cross-linking of their T-cell receptors with anti-TCR mAb or peptide-pulsed antigen-presenting cells. There is a strong correlation between positive selection of thymocytes and CD53 expression. Its association with CD2 and a tyrosine phosphatase in rat T lymphocytes and with CD19, CD21, HLA-DR, and other TM4SF proteins in human B lymphocytes suggests that CD53 is involved in leukocyte signal transduction.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.