Product Information

Description: Recombinant Rhesus Interferon Alpha 2 (Rhesus/Cyno IFN-α2)

Contents: ≥1 x 10⁵ units / vial (MDBK/VSV)

Volume: 0.1 ml

Concentration: ≥ 1 x 10⁶ units/ml (MDBK/VSV)

Buffer: Phosphate buffered saline (PBS) containing 0.1% bovine serum albumin (BSA)

Endotoxin: < 1 EU/µg

Molecular Weight: 19.5 kDa

Purity: ≥ 98%

Source: Rhesus cDNA expressed in E. coli

Accession Number: XM_001107516

Assays Used to Measure Bioactivity: Interferon was titrated in the Cytopathic Effect Inhibition (CPE) assays as listed. The protein has similar activity to Human IFN alpha 2 on human, rhesus and bovine cells.
Bovine (MDBK/VSV)- performed as described [Rubinstein et al. (1981) J. Virol. 37(2):755] The EC₅₀ for interferon in this assay is ~5 U/ml. Specific Activity: (refer to lot specific C.O.A) x 10⁸ units/mg.
Human (A549/EMCV)- performed as described [Budd et al. (1985) Canc. Chem. Pharm. 12:39]. The EC₅₀ for interferon in this assay is ~1 U/ml. Specific Activity: (refer to lot specific C.O.A)x 10⁸ units/mg.
Rhesus (LLC-MK2/VSV)- developed and performed at PBL Biomedical Laboratories. The EC₅₀ for interferon in this assay is ~200 U/ml. Specific Activity: (refer to lot specific C.O.A) x 10⁸ units/mg
As there is no international Monkey Interferon Alpha standard, the units are determined by use of a Human IFN Alpha A (Hu IFN-aA [2a]) laboratory standard calibrated to the international reference standard for Human Interferon Alpha A (Hu IFN-aA [2a]) provided by the National Institutes of Health [Meager et al. (2001). J Immunol Methods. 257(1-2):17-33.].

Shipping Information

Shipping Conditions: Dry ice

Physical State of Product During Shipping: Frozen

Special Conditions/Comments: After receipt, this product should be kept at -70°C or below for retention of full activity. Thaw product vial by incubation in cold tap water until just thawed - the contents of the tube should be apportioned in separate tubes so that freezing and thawing is kept to a minimum. Refreezing should be done on dry ice or in a dry ice/alcohol bath. Further dilution of the product should be in buffers containing protein, such as 0.1% bovine serum albumin (BSA). For more information on protein handling visit the PBL website at www.interferonsource.com.

Product Information: The cDNA for Rhesus/Cyno IFN-α2 was cloned from the Rhesus cell line LLC-MK2. This was found to be identical to the putative Rhesus IFN-α2 (XM_001107516) identified as part of the Rhesus genome project (http://www.hgsc.bcm.tmc.edu/projects/rmacaque). There are 14 AA differences in the mature protein (2 in the signal peptide) between the Rhesus and Human Alpha A (2a) sequence. There are 13 AA differences in the mature protein (2 in the signal peptide) between the Rhesus and Human Alpha 2 (2b) sequence. This is in accord with a previous report [Villinger et al. (1995) J. Immunol. 155:3946] stating that there are 15-16 amino acids different between the Rhesus IFN alpha 2 and human alpha 2 (2b) and human alpha A(2a) respectively.
This Rhesus (Macaca mulatta) sequence is also identical to a Cynomologous (Macaca fascicularis) IFN-α2 cloned from cDNA isolated from primary hepatocytes (PBL unpublished observation).

Comparison of the Mature Sequences of Human and Rhesus IFN Alpha-2

Rhesus   1 CDLPQTHSLG NRRTLMLLAQ MRRISLFFCL KDRHDFEFPQ EEFGNQFQKA QTIPVLHEMI

 Human   1 .......... S........ ........S. .......G... .......... E.........

Rhesus  61 QQTFNLFSTK DSSAAWDETL LNKFYTELYQ QLNDLEACVM QEMGVTETPL MNKNSILAVR

 Human  61 ..I....... .......... .D........ .........I .GV....... .KED......

Rhesus 121 KYFQRITLYL KEKKYSLCAW EVVRAEIMRS FSLSTNLQES LRSKE

 Human 121 .......... ......P... .......... .......... .....

Figure 1: Activity of Rhesus/Cyno and Human IFN Alpha A on rhesus cells (LLC-MK2) challenged with VSV.

Figure 2: Activity of Rhesus/Cyno and Human IFN Alpha A on human cells (A549) challenged with EMCV.

This Product is for RESEARCH USE ONLY and is not for sale or use in any commercial kit or for use in the preparation of any commercial antibody. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE, OR USE IN HUMANS.

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