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苏州工业园区生物纳米园A4#216
联系电话
4000-520-616 / 18915418616
传真号码
0512-67156496
电子邮箱
info@ebiomall.com
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https://www.ebiomall.com
商品描述
Antigen Information
Accession Number:
- Q9BZW8
Gene ID:
- 51744
Gene Symbols:
- 2B4
- CD244
Protein Name & Synonyms:
Natural killer cell receptor 2B4 (NK cell activation-inducing ligand) (NAIL) (NK cell type I receptor protein 2B4) (NKR2B4) (h2B4) (SLAM family member 4) (SLAMF4) (Signaling lymphocytic activation molecule 4) (CD antigen CD244)
Target Species:
- Human
Assay Format
Sensitivity:
0.005 ng/ml (Anticipated minimum sensitivity. Assay may detect lower levels of antigen.)
Number of Targets Detected:
1
Species Detected:
- Human
Compatible Sample Types:
- Cell Culture Supernatants
- Plasma
- Serum
Design Principle:
- Sandwich-based
Method of Detection:
qPCR
Quantitative/Semi-Quantitative:
- Quantitative
Solid Support:
96-well Microplate
Product Specifications
Size:
1, 2, or 5 x 96-Well Strip Microplate Kit
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Introduction
The RayBio® Immuno Quantitative Enzyme Linked ImumunoSorbent Assay (IQELISA™) is an innovative new assay that combines the specificity and ease of use of an ELISA with the sensitivity of real-time PCR. This results in an assay that is simultaneously familiar and cutting edge and enables the use of lower sample volumes while also providing more sensitivity. The RayBio® Human 2B4 IQELISA™ Kit is a modified ELISA assay with high sensitivity qPCR readout for the quantitative measurement of Human 2B4 in serum, plasma, and cell culture supernatants. This assay employs an antibody specific for Human 2B4 coated on a 96-well PCR plate. Standards and samples are pipetted into the wells and 2B4 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and a detection affinity molecule is added to the plates. After washing away unbound detection affinity molecule, primers and a PCR master mix are added to the wells and data is collected using qPCR. Ct values obtained from the qPCR are then used to calculate the amount of antigen contained in each sample, where lower Ct values indicate a higher concentration of antigen.
Typical Data
Application Notes
Kit Components
- 2B4 Microplate (Item A): 96 well PCR plate coated with anti-Human 2B4
- Wash Buffer I Concentrate (20x) (Item B): 25 ml of 20x concentrated solution
- Standards (Item C): 2 vials of recombinant Human 2B4
- Assay Diluent A (Item D): 30 ml diluent buffer, 0.09% sodium azide as preservative. For Standard/Sample (serum/plasma) diluent
- Assay Diluent B (Item E): 15 ml of 5x concentrated buffer. For Standard/Sample (cell culture medium/urine) diluent
- Detection Affinity Reagent for 2B4 (Item F): 2 vials of a 4x concentrated solution of anti-Human 2B4 affinity reagent
- IQELISA™ Detection Reagent (Item G): 1.4ml of a 10x concentrated stock
- Primer Solution (Item I): 1.7 ml vial
- PCR Master Mix (Item J): 1.2 ml vial
- PCR Preparation buffer (Item K): 1ml vial of 10x concentrated buffer
- Final Wash Buffer (Item L): 10ml vial of 10x concentrated buffer
Other Materials Required
- Real-time PCR instrument, Bio-Rad recommended
- 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
- Log-log graph paper or computer and software for data analysis
- Tubes to prepare standard or sample dilutions
- Heating block or water bath capable of 80°C
Protocol Outline
- Prepare all reagents, samples and standards as instructed
- Add 25µl standard or sample to each well. Incubate for 2.5 hours at room temperature or overnight at 4°C
- Add 25 µl detection affinity reagent to each well. Incubate 1 hour at room temperature
- Add 25µL of IQELISA™ Detection Reagent to each well. Incubate 1 hour
- Add 15µL Primer solution 10µL of PCR master mix to each well
- Run real-time PCR
Storage/Stability
May be stored for up to 6 months at 2° to 8°C from the date of shipment. Standard (recombinant protein) should be stored at -20°C or -80°C (recommended at -80°C) after reconstitution. Opened PCR plate or reagents may be stored for up to 1 month at 2° to 8°C. Note: the kit can be used within one year if the whole kit is stored at -20°C. Avoid repeated freeze-thaw cycles.
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