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Boster's ELISA Kit is for the detection of human Alpha B Crystallin in cell lysates, tissue extracts, and serum samples. Each kit contains sufficient components to quantitate the Alpha B Crystallin concentration in up to 40 samples, tested in duplicate.
| Product Name | Alpha B Crystallin ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7105 |
| Description | Colorimetric detection of Alpha B Crystallin. 96wells/kit, with removable strips. |
| Cite This Product | Alpha B Crystallin ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7105) |
| Validated Species | Human |
| Application | ELISA *Our Boster Guarantee covers the use of this product in the above tested applications. **For positive and negative control design, consult "Tissue specificity" under Protein Target Info. |
| Cross Reactivity | There is no detectable cross-reactivity. |
| Pack Size | 96wells/kit, with removable strips. |
| Sensitivity | 0.009 ng/ml *Sensitivity, or Lower Limit of Detection (LLD), is the minimum level of target protein the ELISA assay can detect. We measure 20 blank wells and if the O.D. value is 2 standard deviations higher than the blanks' average O.D. the sample can be deemed positive. |
|---|---|
| Assay Range | 0.352 - 22.5 ng/ml *This assay range is determined using common samples. For samples with low target protein concentrations, users can adjust the standard curve to extend the lower limit of assay range. |
| Sample Type | Cell lysates, Serum, Tissue *The above listed samples are the ones valided with the assay. If you do not see your sample of interest listed, as long as there is enough level of target protein present in the sample, this Picokine? ELISA kit should detect it. **For protocol and tips regarding preparing your sample of interest, please check our ELISA sample preparation guide. |
| Storage | Store at 4°C. |
| Description | Quantity |
| Anti-Alpha B Crystallin Immunoassay Plate | 1 Plate |
| 5X Alpha B Crystallin Extraction Reagent | 1 vial/10 ml |
| Recombinant Alpha B Crystallin Standard | 2 vials |
| Standard and Sample Diluent | 1 vial/ 50 ml |
| 10X Wash Buffer Concentrate | 1 vial/100 ml |
| Anti-Alpha B Crystallin Biotinylated Antibody Concentrate | 1 vial/150 ?l |
| Anti-Alpha B Crystallin Biotinylated Antibody Diluent | 1 vial/ 13 ml |
| Streptavidin: HRP Concentrate | 1 vial/150 ?l |
| Streptavidin: HRP Diluent | 1 vial/ 13 ml |
| TMB Substrate | 1 vial/ 13 ml |
| Stop Solution | 1 vial/ 13 ml |
1. Ultra pure water.
2. Additional reagents and materials for cell lysate and tissue extract preparation, including protease inhibitors.
3. Precision pipettors, with disposable plastic tips.
4. Polypropylene or polyethylene tubes to prepare samples ? do not use polystyrene, polycarbonate or glass tubes.
5. A container to prepare 1X Wash Buffer.
6. A wash bottle or an automated 96-well plate washer.
7. Disposable reagent reservoirs.
8. A standard microtiter plate reader for measuring absorbance at 450 nm.
9. Adhesive plate sealers.
You can check the tissue specificity below for information on selecting positive and negative control.
| Gene Name | CRYAB |
|---|---|
| Protein Name | Alpha-crystallin B chain |
| Protein Function | May contribute to the transparency and refractive index of the lens. Has chaperone-like activity, preventing aggregation of various proteins under a wide range of stress conditions. |
| Tissue Specificity | Lens as well as other tissues (PubMed:838078, PubMed:2387586). Expressed in myocardial tissue (PubMed:28493373). |
| Subcellular Localization | Cytoplasm. |
| Uniprot ID | P02511 |
| Alternative Names | Alpha-crystallin B chain; Alpha(B)-crystallin; Heat shock protein beta-5; HspB5; Renal carcinoma antigen NY-REN-27; Rosenthal fiber component; CRYAB; CRYA2; HSPB5 |
| Research Areas | Cancer, Heat Shock| |
*if product is indicated to react with multiple species, protein info is based on the human gene.
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 . 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.
Click the images to enlarge.
Typical Standard Curve for the Alpha B Crystallin ELISA Kit (Enzyme-Linked Immunosorbent Assay) - EK7105. Assay Type: Sandwich ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.352 - 22.5 ng/mL.
1. Prepare Standard and samples in Standard and Sample Diluent.
2. Add 100 ?L 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 ?L 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 ?L 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 ?L 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 ?L of Stop Solution to each well.
14. Measure absorbance on a plate reader at 450 nm.
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