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Rat PR domain zinc finger protein 16 (PRDM16) ELISA Kit


Species Reactivity

Rat (Rattus norvegicus)

UniProt

N/A

Abbreviation

PRDM16

Alternative Names

KIAA1675; MEL1; MGC166915; PFM13; MDS1/EVI1-like|PR-domain zinc finger protein 16|transcription factor MEL1

Range

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Sensitivity

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Sample Type

Serum, Plasma, Other biological fluids

Detection Method

Sandwich

Analysis Method

Quantitive

Assay Duration

1-4.5h

Sample Volume

1-200 μL

Detection Wavelengt

450 nm

Cat

AE64512RA

Size

48T 96T

Price

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Availability

3-5 working days

Manual

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Test principle:
This assay employs a two-site sandwich ELISA to quantitate PRDM16 in samples. An antibody specific for PRDM16 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any PRDM16 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PRDM16 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of PRDM16 bound in the initial step. The color development is stopped and the intensity of the color is measured.

Product Overview:
The 1,257-amino acid MEL1 protein shares 56% amino acid identity with MDS1/EVI1 and has a similar domain structure. The first 222 amino acids of MEL1 are highly homologous to the PR domain of MDS1, and the rest of the MEL1 sequence is homologous to EVI1. Following the N-terminal PR domain, MEL1 has a zinc finger DNA-binding domain, a proline-rich domain, a repressor domain, a second zinc finger DNA-binding domain, and a C-terminal acidic domain. Northern blot and RT-PCR analyses of various leukemia cell lines revealed a major 8-kb MEL1 transcript only in cells with t(1;3). In human tissues, RT-PCR showed MEL1 expression only in uterus and fetal kidney. MEL1 and MEL1S had apparent molecular masses of 170 and 150 kD, respectively, and t(1;3)-positive leukemias expressed predominantly MEL1S protein.

Components:

Reagents

Quantity

Reagents

Quantity

Assay plate (96 Wells)

1

Instruction manual

1

Standard (lyophilized)

2

Sample Diluent

1 x 20 mL

Biotin-Conjugate (concentrate 100 x)

1 x 120 μL

Biotin-Conjugate Diluent

1 x 20 mL

Streptavidin-HRP  (concentrate 100 x)

1 x 120 μL

Streptavidin-HRP Diluent   

1 x 20 mL

Wash Buffer (concentrate 25 x)

1 x 20 mL

Substrate Solution

1 x 12 mL

Stop Solution

1 x 10 mL

Adhesive Films

4

Specificity:

This assay has high sensitivity and excellent specificity for detection of Rat PRDM16. No significant cross-reactivity or interference between Rat PRDM16 and analogues was observed.


Recovery:
Matrices listed below were spiked with certain level of recombinant Rat PRDM16 and the recovery rates were calculated by comparing the measured value to the expected amount of Rat PRDM16 in samples.

Sample Type

Number

Recovery range (%)

Average(%)

Serum

10

90-101

96

EDTA plasma

10

89-97

93

Heparin plasma

10

91-99

95

Precision:
Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%

Linearity:
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat PRDM16 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample Type

1:2

1:4

1:8

1:16

Serum

78-89%

81-99%

92-103%

95-105%

EDTA plasma

91-101%

90-98%

93-101%

91-98%

Heparin plasma

92-103%

93-102%

92-99%

91-101%

Stability:
The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).

Sample collection and storage:
Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.

Kits storage instructions:
Store at 2-8°C. Please refer to Instruction Manual.