Various Species ARG ELISA Kit, Lot 21AO-1041 [Cancer Immune Checkpoint Assay Kit] (CAT#: IOK-05-P952)

Assay plate x1
Standard x2
Sample Diluent 1 x 20ml
Assay Diluent A 1 x 10ml
Assay Diluent B 1 x 10ml
Detection Reagent A 1 x 120μl
Detection Reagent B 1 x 120μl
Wash Buffer(25 x concentrate) 1 x 30ml
Substrate 1 x 10ml
Stop Solution 1 x 10ml
Plate sealer for 96 wells x5
Instruction 1x

Microplate reader.
Pipettes and pipette tips.
EP tube Deionized or distilled water.

50 μL

3 - 5 h

Pre-coated

Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix gently until the crystals have completely dissolved. Dilute 30 mL of Wash Buffer Concentrate into deionized or distilled water to prepare 750 mL of Wash Buffer.
Standard - Reconstitute the Standard with 1.0 mL of Sample Diluent. This reconstitution produces a stock solution of 200 umol/L. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making serial dilution. The undiluted standard serves as the high standard (200 umol/L). The Sample Diluent serves as the zero standard (0 umol/L).

1.Add 100µL of standard or sample per well. Cover with the adhesive strip provided. Incubate for 2 hours at 37°C.
2.Remove the liquid of each well, don't wash.
3.Add 100µL of Biotin-antibody (1×) to each well. Cover with a new adhesive strip. Incubate for 1 hour at 37°C.
4.Aspirate each well and wash, repeating the process two times for a total of three washes.After the last wash, remove any remaining wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
5.Add 100µL of HRP-avidin (1×) to each well. Cover the microtiter plate with a new adhesive strip. Incubate for 1 hour at 37°C.
6.Repeat the aspiration/wash process for five times as in step 6.
7.Add 90µL of TMB Substrate to each well. Incubate for 20 minutes at 37°C. Protect from light.
8.Add 50µL of Stop Solution to each well, gently tap the plate to ensure thorough mixing.
9.Determine the optical density of each well within 5 minutes using a microplate reader set to 450nm.

Average the duplicate readings for each standard, control, and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit.

The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

4 °C

The Assay Plate, Standard, Detection Reagent A and Detection Reagent B should be stored at -20°C upon being received. After receiving the kit , Substrate should be always stored at 4°C.Other reagents are kept according to the labels on vials. But for long term storage, please keep the whole kit at -20°C.

6 months

The Assay Plate, Standard, Detection Reagent A and Detection Reagent B should be stored at -20°C upon being received. After receiving the kit , Substrate should be always stored at 4°C.Other reagents are kept according to the labels on vials. But for long term storage, please keep the whole kit at -20°C.

For Research Use only

Arginase

CG18104; Dmel\\CG18104; EG:171D11.4; EG:65F1.3; SI:zC146F4.4 (novel protein with NUDIX domain); si:ch211-146f4.3; Tb08.26N11.490; rocF; NV10213; argi1; arginase; arginase 1 L homeolog; arginase 1; Arginase-1; arg; arg1.L; arg1; ARGAH1; BP0538; rocF; SAS2066; RR_RS05730; CND03500; CNG00550; Tb927.8.2020; PGTG_16455; LOC100123155; argi1

The microtiter plate provided in this kit has been pre-coated with an antibody specific to Arginine. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for Arginine and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm.