Protein Name & Synonyms:
Zinc-alpha-2-glycoprotein (Zn-alpha-2-GP) (Zn-alpha-2-glycoprotein)
Recommended Dilution (serum & plasma):
Human: 4X / Mouse: 2X / Rat: 10X
This kit detects ZAG 278aa. No other active isoforms have been reported.
Cross Reactivity: This EIA kit shows no cross-reactivity with any of the cytokines tested: Ghrelin, Nesfatin, NPY and APC.
Number of Targets Detected:
Compatible Sample Types:
- Cell Culture Supernatants
Method of Detection:
1, 2, or 5 x 96-Well Strip Microplate Kit
ZAG is a 41 kDa soluble glycoprotein, which is a homologue of the class 1 major histocompatibility complex (MHC) heavy chain. ZAG was found to stimulate lipolysis through stimulation of adenylyl cyclase in a GTP-dependent process. ZAG produced weight loss when administered to obese mice without a reduction in food and water intake. Loss of body weight was entirely due to loss of adipose tissue with no effect on lean body mass. There was a three-fold increase in oxygen consumption by interscapular brown adipose tissue (BAT), suggesting an increase in thermogenesis. Recent studies have reported that ZAG is not only capable of stimulating lipolysis through activation of HSL, but also increases the utilization of the released NEFA through an increased level of UCPs in BAT and skeletal muscle.
- Strip plates and additional reagents allow for use in multiple experiments
- Quantitative protein detection
- Establishes normal range
- The best products for confirmation of antibody array data
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Standard Peptide
- Assay Diluent(s)
- Biotinylated Peptide
- TMB One-Step Substrate
- Stop Solution
- Assay Diagram
- Positive Control Sample
- Capture Antibody
- User Manual
Other Materials Required
- Distilled or deionized water
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare standard and sample dilutions
- Orbital shaker
- Aluminum foil
- Saran Wrap
- Absorbent paper
- Microplate reader capable of measuring absorbance at 450nm
- SigmaPlot software (or other software that can perform four-parameter logistic regression models)
- Prepare all reagents, samples and standards as instructed.
- Add 100 µl detection antibody to each well.
- Incubate 1.5 h at RT or O/N at 4°C.
- Add 100 µl standard or sample to each well.
- Incubate 2.5 h at RT.
- Add 100 µl prepared streptavidin solution.
- Incubate 45 min at RT.
- Add 100 µl TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl Stop Solution to each well.
- Read plate at 450 nm immediately.
Standard, Biotinylated Zinc alpha 2-glycoprotein (ZAG) peptide, and Positive Control should be stored at -20°C after arrival. Avoid multiple freeze-thaws. The remaining kit components may be stored at 4°C. Opened Microplate Wells and antibody (Item N) may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge.