Human Vitamin B12,VB12 ELISA Kit

Instructions
Code CSB-E07903h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human Vitamin B12 (VB12) ELISA Kit is applied to detect and quantify the concentrations of VB12 in serum and plasma. This kit exclusively recognizes human VB12. It adopts the Competitive ELISA technique in which Biotin-conjugated VB12 and VB12 in samples or standards compete for binding to the pre-coated anti-human VB12 antibody. After washing, HRP-avidin is added to the wells. And the chromogenic reaction is triggered after the addition of TMB substrate solution. After adding the stop solution, the color development is immediately terminated and the color turns from blue to yellow. The intensity of the color is negatively relevant to the levels of VB12 in the sample. This ELISA kit has been confirmed to have high sensitivity, excellent specificity, premium precision, high recovery, and lot-to-lot consistency. See the product instructions for more details.

Vitamin B12, also named cobalamin, is a water-soluble vitamin naturally available in all animal tissues. It plays crucial roles in proper red blood cell formation, neurological function, and DNA synthesis. Vitamin B12 also acts as a coenzyme that is essential for the enzymatic catalytic activities of methionine synthase and L-methylmalonyl-CoA mutase. Methionine synthase catalyzes the conversion of homocysteine to methionine, which is required for the formation of S-adenosylmethionine that donates the methyl group for many substrates, such as DNA, RNA, and proteins. The conversion of L-methylmalonyl-CoA to succinyl-CoA is an essential biochemical reaction in fat and protein metabolism during propionate degradation. Succinyl-CoA is also required for hemoglobin synthesis. We can obtain vitamin B12 from various animal-derived foods and supplement form. Deficiency of Vitamin B12 can lead to irreversible neurological difficulties and anemia.

Target Name Vitamin B12,VB12
Abbreviation VB12
Species Homo sapiens (Human)
Sample Types serum, plasma
Detection Range 1.56 pg/mL-100 pg/mL
Sensitivity 1.498 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Metabolism
Assay Principle quantitative
Measurement Competitive
Precision

 

Intra-assay Precision (Precision within an assay): CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess.

Inter-assay Precision (Precision between assays): CV%<10%

Three samples of known concentration were tested in twenty assays to assess.

 

Linearity

 

To assess the linearity of the assay, samples were spiked with high concentrations of human VB12 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.

 

Typical Data

 

These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.

 

Materials provided
  • A 96-well Assay plate --The 96-well plate has been pre-coated with anti-human VB12 antibody.
  • Standard (2 x 0.5 ml)--Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
  • Biotin-conjugated VB12(100 x concentrate) (1 x 600 μl)
  • HRP-avidin (100 x concentrate) (1 x 120 μl)
  • Biotin-conjugate Diluent (1 x 10 ml) --Dilute the Biotin-conjugated VB12
  • HRP-avidin Diluent (1 x 20 ml) --Dilute the HRP-avidin solution.
  • Sample Diluent (2 x 20 ml) --Reconstitute the standard and dilute the sample to an appropriate concentration.
  • Wash Buffer (25x concentrate) (1 x 20 ml)--Wash away unbound or free substances.
  • TMB Substrate (1x 10 ml) --Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • Stop Solution (1 x 10ml) --Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells)
  • An Instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm - 570 nm.
  • An incubator that can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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 Q&A
Q:

Protocol P9, step 3 - Set a Blank well without any solution.
This is a completely empty well with no solution or liquid at all? Am I correct?
May I know if this is S0. If it is S0, do we add the wash buffer, TMB and stop solution?

A:
Thanks for your inquiry.
Actually step3 is just only one procedure during the whole experiment. You need to check the full steps according our manual.
This kit is competitive one and the blank is different from S0 indeed. The blank should be added Wash Buffer+TMB Substrate+Stop Solution.
You could see it from the protocol. Pls read the manual carefully.

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