CRYAB ELISA Kit

Catalog No. ABIN2964835

This Colorimetric ELISA kit is designed for the quantitative measurement of Human CRYAB. There is 1 publication available.

Quick Overview for CRYAB ELISA Kit (ABIN2964835)

Target: CRYAB (Crystallin, alpha B (CRYAB))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.352 ng/mL - 22.5 ng/mL

Application: ELISA

Sample Type: Cell Lysate, Tissue Samples, Serum

Product Details CRYAB ELISA Kit

Minimum Detection Limit: 0.352 ng/mL

Purpose: Colorimetric detection of alpha B Crystallin

Analytical Method: Quantitative

Sensitivity: 0.009 ng/mL

Characteristics: ELISA kit used to quantitate alpha B crystallin concentration in samples.

Components:

• Anti-Alpha B Crystallin Immunoassay Plate
• 5X Alpha B Crystallin Extraction Reagent
• Recombinant Alpha B Crystallin Standard
• Standard and Sample Diluent
• 10X Wash Buffer Concentrate
• Anti-Alpha B Crystallin Biotinylated Antibody Concentrate
• Anti-Alpha B Crystallin Biotinylated Antibody Diluent
• Streptavidin: HRP Concentrate
• Streptavidin: HRP Diluent
• TMB Substrate
• Stop Solution

Material not included: - Ultra pure water
- Additional reagents and materials for cell lysate and tissue extract preparation, including protease inhibitors
- Precision pipettors, with disposable plastic tips
- Polypropylene or polyethylene tubes to prepare samples − do not use polystyrene, polycarbonate or glass tubes
- A container to prepare 1X Wash Buffer
- A wash bottle or an automated 96-well plate washer

Application Details

Assay Time: 0.5 h

Plate: Pre-coated

Protocol:

1. Prepare Standard and samples in Standard and Sample Diluent.
2. Add 100 μL of Standard or sample to appropriate wells.
3. Cover plate with Plate Sealer and incubate at room temperature (20-25 °C) for 1 hour.
4. Wash plate four times with 1X Wash Buffer.
5. Add 100 μL of Biotinylated Antibody Working Solution to each well.
6. Cover plate with Plate Sealer and incubate at room temperature for 1 hour.
7. Wash plate four times with 1X Wash Buffer.
8. Add 100 μL of Streptavidin-HRP Working Solution to each well.
9. Cover plate with Plate Sealer and incubate at room temperature for 30 minutes.
10. Wash plate four times with 1X Wash Buffer.
11. Add 100 μL of TMB Substrate to each well.
12. Develop the plate in the dark at room temperature for 30 minutes.
13. Stop reaction by adding 100 μL of Stop Solution to each well.
14. Measure absorbance on a plate reader at 450 nm.

Assay Procedure:

1. Prepare Standard and samples in Standard and Sample Diluent.
2. Add 100 μL of Standard or sample to appropriate wells.
3. Cover plate with Plate Sealer and incubate at room temperature (20-25 °C) for 1 hour.
4. Wash plate four times with 1X Wash Buffer.
5. Add 100 μL of Biotinylated Antibody Working Solution to each well.
6. Cover plate with Plate Sealer and incubate at room temperature for 1 hour.
7. Wash plate four times with 1X Wash Buffer.
8. Add 100 μL of Streptavidin-HRP Working Solution to each well.
9. Cover plate with Plate Sealer and incubate at room temperature for 30 minutes.
10. Wash plate four times with 1X Wash Buffer.
11. Add 100 μL of TMB Substrate to each well.
12. Develop the plate in the dark at room temperature for 30 minutes.
13. Stop reaction by adding 100 μL of Stop Solution to each well.
14. Measure absorbance on a plate reader at 450 nm.

Calculation of Results:

Duplicate absorbance values should be within 10% of each other. Care should be taken when interpreting data with differences in absorbance values greater than 10%.

1. Prepare a standard curve to determine the amount of Alpha B Crystallin in an unknown sample. Plot the average absorbance obtained for each standard concentration on the vertical (Y) axis versus the corresponding Alpha B Crystallin concentration on the horizontal (X) axis using graph paper or curve-fitting software.

2. Calculate the Alpha B Crystallin concentration in unknown samples using the prepared standard curve. Determine the amount of Alpha B Crystallin in each unknown sample by noting the Alpha B Crystallin concentration (X axis) that correlates with the absorbance value (Y axis) obtained for the unknown sample.

3. Multiply the Alpha B Crystallin concentration obtained by the dilution factor to determine the amount of Alpha B Crystallin in the undiluted sample.

Restrictions: For Research Use only

Handling

Storage: 4 °C

References for CRYAB ELISA Kit (ABIN2964835)

Baba, Oshitari, Yamamoto: "Level of vitreous alpha-B crystallin in eyes with rhegmatogenous retinal detachment." in: Graefe's archive for clinical and experimental ophthalmology, Vol. 253, Issue 8, pp. 1251-4, (2015) (PubMed).

Target Details for CRYAB

Target: CRYAB (Crystallin, alpha B (CRYAB))

Alternative Name: alpha B Crystallin

Background: The alpha-crystallins are major water-soluble lens structural proteins of the vertebrate eye that are related to the small heat shock protein family. The alpha-crystallins possess structural and functional similarities with HSP25 and HSP27. Mammalian lens cystallins are divided into alpha, beta and gamma families. alpha and beta families are further divided into acidic and basic groups (alpha-A and alpha-B respectively). In the lens, alpha-crystallin primarily functions to maintain proper refractive index, however it can also function as a molecular chaperone that binds to the denatured proteins, keeping them in solution and thereby maintaining the translucency of the lens. When cellular stress occurs, alpha-crystallin enters its' phosphorylated state and may serve a structural control function and play a role in protein maintenance. In addition to their interaction with proteins, alpha-crystallins also interact with native molecules such as membrane proteins, Golgi matrix protein, structural proteins, nuclear proteins and DNA (3, 4, 5, 6, and 7). Two other functions are an autokinase activity and participation in the intracellular architecture, and it has also been proven that both alpha-A and B prevent apoptosis by inhibiting caspases. Specifically, alpha-B cystallin is found in many cells and organs outside the lens, and alpha B is overexpressed in several neurological disorders and in cell lines under stress conditions.