5-Hydroxymethylcytosine ELISA Kit

Catalog No. ABIN8067734

Cow, Horse, Human, Mouse, Pig, Rat and Sheep 5-Hydroxymethylcytosine ELISA Kit, Colorimetric assay for quantification of Cow, Horse, Human, Mouse, Pig, Rat and Sheep 5-Hydroxymethylcytosine.

Quick Overview for 5-Hydroxymethylcytosine ELISA Kit (ABIN8067734)

Target: 5-Hydroxymethylcytosine (5-hmC)

Reactivity: Cow, Horse, Human, Mouse, Pig, Rat, Sheep

Detection Method: Colorimetric

Method Type: Competition ELISA

Detection Range: 0.1 pg/mL - 100000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Plasma, Serum, Urine

Product Details 5-Hydroxymethylcytosine ELISA Kit

Minimum Detection Limit: 0.1 pg/mL

Purpose: 5-hmC ELISA Kit is an in vitro quantitative assay for detecting 5-Hydroxymethylation based on the competitive enzyme immunoassay principle.

Analytical Method: Quantitative

Specificity: This EIA kit detects 5-hmC / 5-Hydroxymethylcytosine

Sensitivity: 1.006 pg/mL

Components:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Lyophilized Standard
• Capture Antibody
• Assay Diluent
• Biotinylated Peptide
• HRP-Streptavidin
• Positive Control
• TMB One-Step Substrate Reagent
• Stop Solution

Material not included:

• Microplate reader capable of measuring absorbance at 450 nm
• 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
• Absorbent paper
• Distilled or deionized water
• Software which can perform four-parameter logistic regression models
• Tubes to prepare standard or sample dilutions
• Orbital shaker
• Aluminum foil
• Plastic wrap

Application Details

Application Notes: Suggested Plasma Dilution: 2-4 folds

Sample Volume: 100 μL

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards as instructed.
2. Add 100 μL Capture Antibody to each well. Incubate 1.5 hours at room temperature or overnight at 4 °C.
3. Add 100 μL standard or sample to each well. Incubate 2.5 hours at room temperature or overnight at 4 °C.
4. Add 100 μL prepared Streptavidin solution. Incubate 45 minutes at room temperature.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: The entire kit may be stored at -20°C to -80°C for up to 1 year from the date of shipment. For extended storage, it is recommended to store it at -80°C. Avoid repeated freeze-thaw cycles.

Target Details for 5-Hydroxymethylcytosine

Target: 5-Hydroxymethylcytosine (5-hmC)

Alternative Name: 5-hmC

Target Type: Chemical

Background: Optional[Gene Symbols]: 5-hydroxymethylcytosine (5-hmC), also known as the ""sixth base"" of DNA, is generated by the oxidation of 5-methylcytosine (5-mC) through the action of the ten-eleven translocation (TET) family of enzymes. 5-hmC is thought to be the DNA demethylation pathway, thereby playing a crucial part in gene regulation, chromatin structure and accessibility, recruiting or repelling transcription factors, and ultimately modulating gene expression. The presence of 5-hmC is particularly abundant in neuronal cells, suggesting a significant role in brain development and function. In addition, levels of 5-hmC undergo highly dynamic changes during development, differentiation and cancer. 5-hmC plays a critical role in developmental processes and dysregulation of 5-hmC has been associated with several human diseases.