Chicken HMGB1 ELISA Kit, Lot 21AO-434 [Cancer Immune Checkpoint Assay Kit]

The kit is designed for in vitro quantitative measurement of Chicken HMGB1 in Plasma, Serum, Tissue Homogenate.

This immunoassay kit allows for the in vitro quantitative determination of chicken high mobility group protein b1,High mobility group protein B1 concentrations in serum, Plasma, tissue homogenates and Cell culture supernates and Other biological fluids.

ELISA

The kit should not be used beyond the expiration date on the kit label.
Do not mix or substitute reagents with those from other lots or sources.
If samples generate values higher than the highest standard, further dilute the samples with the Sample Diluent and repeat the assay. Any variation in standard diluent, operator, pipetting technique, washing technique,incubation time or temperature, and kit age can cause variation in binding.

HMGB1

Chicken

Colorimetric

Sandwich ELISA

Quantitative

Plasma, Serum, Tissue Homogenate

This assay recognizes recombinant and natural chicken High mobility group protein B1. No significant cross-reactivity or interference was observed.
Note: Limited by current skills and knowledge, it is impossible for us to complete the cross- reactivity detection between chicken High mobility group protein B1 and all the analogues, therefore, cross reaction may still exist.

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.

100 μL

3 - 5 h

Pre-coated

Wash Buffer -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. The undiluted standard serves as the high standard (10.0 ng/mL). The Sample Diluent serves as the zero standard (0 ng/mL).
Standard - Reconstitute the Standard with 1.0 mL of Sample Diluent. This reconstitution produces a stock solution of 10.0 ng/mL. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making serial dilutions (Making serial dilution in the wells directly is not permitted). The undiluted standard serves as the high standard (10.0 ng/mL). The Sample Diluent serves as the zero standard (0 ng/mL).

1.Add 100 μL of Standard, Blank, or Sample per well. Cover with the Plate sealer. Incubate for 2 hours at 37 °C.
2.Remove the liquid of each well, don't wash. Add 100 μL of Detection Reagent A working solution to each well. Incubate for 1 hour at 37 °C.
3.Aspirate each well and wash, repeating the process three times for a total of three washes. let it sit for 1~2 minutes. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
4.Add 100 μL of Detection Reagent B working solution to each well. Cover with a new Plate sealer. Incubate for 1 hour at 37 °C.
5.Repeat the aspiration/wash process for 5 times as conducted in step three.
6.Add 90 μL of Substrate Solution to each well. Incubate within 15-30 minutes at 37 °C. Protect from light.
7.Add 50 μL of Stop Solution to each well.
8.Determine the optical density of each well at once, using a microplate reader set to 450 nm.

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. The unused strips should be kept in a sealed bag with the desiccant provided to minimize exposure to damp air. The test kit may be used throughout the expiration date of the kit.

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. The unused strips should be kept in a sealed bag with the desiccant provided to minimize exposure to damp air. The test kit may be used throughout the expiration date of the kit.

For Research Use only

High Mobility Group Protein 1 (HMGB1)

HMG1; HMG3; SBP-1; DEF; HMG-1; Hmg1; amphoterin; p30; hmgb1; ik:tdsubc_1a5; wu:fb23c02; xx:tdsubc_1a5; zgc:56110; zgc:77104; hmg-1; hmg3; sbp-1; hmg1; HMGB1; Ac2-008; high mobility group box 1; high-mobility group box 1; high mobility group box 1a; high mobility group box 1 L homeolog; high mobility group protein B1; HMGB1; Hmgb1; hmgb1; hmgb1a; hmgb1.L; LOC100359149

Synonyms: High mobility group protein B1, High mobility group protein 1(HMG-1)

P36194

p53 Signaling, Regulation of Muscle Cell Differentiation, Skeletal Muscle Fiber Development, Positive Regulation of Endopeptidase Activity, Regulation of Carbohydrate Metabolic Process, Toll-Like Receptors Cascades, Smooth Muscle Cell Migration, Inflammasome

The microtiter plate provided in this kit has been pre-coated with an antibody specific to High mobility group protein B1. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for High mobility group protein B1 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. The concentration of High mobility group protein B1 in the samples is then determined by comparing the O.D. of the samples to the standard curve.