Af498

Manufactured by R&D Systems

AF498 is a laboratory instrument designed for the measurement and analysis of sample materials. It features advanced technology to provide accurate and reliable results. The core function of AF498 is to facilitate the evaluation and characterization of various substances and samples in a controlled laboratory environment.

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Lab products found in correlation

Recombinant mouse leptin, by Merck Group (1 mentions) L1381, by Merck Group (1 mentions)

3 protocols using af498

Neutralizing Leptin's Effects in DIO Mice

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To neutralize leptin’s actions a goat IgG against recombinant mouse leptin (AF498 from R and D systems, Minneapolis) was peripherally (i.p.) administered into DIO mice for 5 consecutive days, following a sham injection period. Mice received i.p. injections of 24 μg in 90 μl of antibody solution or 90 μl of IgG control (Antibodies Australia, Victoria, Australia). The dose of the antibody was determined from previous research (Konstantinides et al., 2004 (link)). All mice where randomly treated.

Simonds S.E., Pryor J.T., Ravussin E., Greenway F.L., Dileone R., Allen A.M., Bassi J., Elmquist J.K., Keogh J.M., Henning E., Myers MG J.r., Licinio J., Brown R.D., Enriori P.J., O’Rahilly S., Sternson S.M., Grove K.L., Spanswick D.C., Farooqi I.S, & Cowley M.A. (2014). Leptin Mediates the Increase in Blood Pressure Associated with Obesity. Cell, 159(6), 1404-1416.

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LPC-Induced Spinal Cord Injury Model

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Adult female mice at 7–8 weeks of age underwent laminectomy at Th11/Th12; 2 μl of 10 mg/ml lysophosphatidylcholine (LPC, L1381, Sigma Aldrich) dissolved in PBS was injected into the midline dorsal column at a depth of 0.5 mm13 (link). For administration of recombinant mouse leptin (Sigma Aldrich) or leptin neutralizing antibodies (AF498, R&D System), a cannula from Alzet osmotic pump (model No. 1002 or 1007D, Alzet Corp.) was placed at the thoracic spinal cord under the dura 3 days after LPC injection. The pump was filled with vehicle solution (PBS); recombinant mouse leptin (12 μg/kg body weight per day) or leptin neutralizing antibodies (10 μg/kg of body weight per day) were administered subcutaneously on the back.

Matoba K., Muramatsu R, & Yamashita T. (2017). Leptin sustains spontaneous remyelination in the adult central nervous system. Scientific Reports, 7, 40397.

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Leptin ELISA Assay Protocol

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ELISA plate was coated with 50uL capture antibody (1:1000 in PBS) and put at 4°C overnight. Next day plate was washed 3 times with Wash Buffer (1L PBS + 0.05% Tween 20). Plate was Blocked with 200uL blocking buffer (200mL PBS + 1% BSA) for 1 hour at room temperature. Samples were added (50ul) in blocking buffer to the wells together with Standard Curve samples. Plate was incubated at room temperature for 2 hours. Secondary antibody was added (1:2000 in blocking buffer) and incubated at room temperature for 1 hour. After one hour HRP streptavidin (1:2000 in blocking buffer) was added and incubated at room temperature for 30min. We added 40uL TMB substrate A and 40uL TMB substrate B to develop samples. Plate was read at 450nm in a plate reader. Antibodies used for leptin Elisa experiment: Capture Mouse Leptin/OB antibody (R&D systems AF498) and detection antibody Mouse Leptin/OB antibody (R&D system BAF498).

Rivadeneira D.B., DePeaux K., Wang Y., Kulkarni A., Tabib T., Menk A.V., Sampath P., Lafyatis R., Ferris R.L., Sarkar S.N., Thorne S.H, & Delgoffe G.M. (2019). Oncolytic viruses engineered to enforce leptin expression reprogram tumor-infiltrating T cell metabolism and promote tumor clearance. Immunity, 51(3), 548-560.e4.

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