Il12a ELISA Kit (Rat) (OKBB01155)

Catalog No: OKBB01155

Size:96 wells

Il12a ELISA Kit (Rat) (OKBB01155) Standard Curve

Product Info

• Predicted Species Reactivity: Rat
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's rat IL-12(P70) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for IL-12(P70) has been precoated onto 96-well plates. Standards (Expression system for standard: NS0, Immunogen sequence: (M23-S335)+(R23-S215)) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for IL-12(P70) is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the rat IL-12(P70) amount of sample captured in plate.
• ELISA Kit Range: 15.6 pg/ml - 1,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  n	16	16	16	24	24	24
  Mean(pg/ml)	35	222	502	34	224	454
  Standard deviation	1.92	15.76	27.61	2.07	19.93	29.05
  CV(%)	5.5	7.1	5.5	6.1	8.9	6.4

• ELISA Kit Component:
  • 96-Well Plate Pre-coated with Anti-Rat Il12a Antibody: 96 Wells (12 x 8 Well Strips)
  • Lyophilized Recombinant Rat Il12a Standard: 2 x 10 ng
  • 100X Biotinylated Anti-Rat Il12a Antibody: 1 x 130 uL
  • 100X Avidin-Biotin Peroxidase Complex (ABC): 1 x 130 uL
  • Sample Diluent Buffer: 1 x 30 mL
  • Antibody Diluent Buffer: 1 x 12 mL
  • ABC Diluent Buffer: 1 x 12 mL
  • TMB Color Developing Agent: 1 x 10 mL
  • TMB Stop Solution: 1 x 10 mL
  • 10X Wash Buffer: 1 x 30 mL
• Additional Information:

  To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.

  Lots	Lot1 (pg/ml)	Lot2 (pg/ml)	Lot3 (pg/ml)	Lot4 (pg/ml)	Mean (pg/ml)	Standard Deviation	CV (%)
  Sample 1	35	31	36	34	34	1.87	5.5%
  Sample 2	222	226	219	229	224	3.8	1.6%
  Sample 3	502	447	478	479	476	19.55	4.1%

  *number of samples for each test n=16.

  
  

  Standard Number	8	7	6	5	4	3	2	1
  Concentration (pg/ml)	0	15.6	31.2	62.5	125	250	500	1000
  O.D.	0.019	0.121	0.214	0.384	0.632	1.020	1.513	2.221

• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates and serum
• Sensitivity: <1.5 pg/ml
• Specificity: Natural and recombinant rat IL-12(P70)
• Immunogen: Expression system for standard: NS0; Immunogen sequence:(M23 - S335) + (R23 - S215)
• Description: For Quantitative Detection of rat IL-12(P70) in cell culture supernatants and serum.

Target Info

• Gene Symbol: IL12A
• Gene Full Name: interleukin 12A
• Alias Symbols: Interleukin-12 subunit alpha; IL-12A; Cytotoxic lymphocyte maturation factor 35 kDa subunit; CLMF p35; IL-12 subunit p35; Il12a
• NCBI Gene Id: 4405
• Protein Name: Interleukin-12 subunit alpha
• Description of Target: Interleukin (IL)-12 is a 70-KDa cytokine comprised of two disulfide-linked proteins (p35 and p40) and is essential for the initiation of effective immune response. And the IL-12p70 is a heterodimer of p35 and p40 subunits; it is an important cytokine secreted by antigen-presenting cells in response to antigenic stimulation. Gene expression analysis of the IL-12 cytokine family subunits revealed that both strains induced high levels of p40 (protein chain communal to IL-12 p70 and IL-23) as well as p19, a subunit of IL-23. Conversely only ACT- 18HS19 infection induced consistent transcription of IL-12 p35, a subunit of IL-12 p70. The standard product used in this kit is recombinant rat IL-12 p70 comprising of a 193 amino acids p35 chain and a 313 amino acids p40 chain connected by a disulfide bond.
• Uniprot ID: Q9R103