HSP90 ELISA Kit

Catalog No. ABIN2964825

Product Details HSP90 ELISA Kit

Target: HSP90 (Heat Shock Protein 90 (HSP90))

Reactivity: Human, Goat, Zebra Finch

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.44 ng/mL - 28 ng/mL

Minimum Detection Limit: 0.44 ng/mL

Application: ELISA

Purpose: Colorimetric detection of HSP90 alpha

Sample Type: Blood, Cell Lysate, Serum, Tissue Samples

Analytical Method: Quantitative

Sensitivity: 0.117 ng/mL

Characteristics: ELISA kit used to quantitate HSP90 alpha concentration in samples.

Components:

• Anti-Hsp90a Immunoassay Plate
• 5X Hsp90a Extraction Reagent
• Recombinant Hsp90a Standard
• Standard and Sample Diluent
• 10X Wash Buffer Concentrate
• Anti-Hsp90a Biotinylated Antibody Concentrate
• Anti-Hsp90a 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 37 °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 37 °C 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 37 °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, 20-25 °C 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 Hsp90α in an unknown sample. Plot the average absorbance obtained for each standard concentration on the vertical (Y) axis versus the corresponding Hsp90α concentration on the horizontal (X) axis using graph paper or curve-fitting software.

2. Calculate the Hsp90α concentration in unknown samples using the prepared standard curve. Determine the amount of Hsp90α in each unknown sample by noting the Hsp90α concentration (X axis) that correlates with the absorbance value (Y axis) obtained for the unknown sample.

3. Multiply the Hsp90α concentration obtained by the dilution factor to determine the amount of Hsp90α in the undiluted sample.

Restrictions: For Research Use only

Handling

Storage: 4 °C

References for HSP90 ELISA Kit (ABIN2964825)

Miyazaki, Nakamura, Hineno, Kobayashi, Kinoshita, Yoshida, Ikeda: "Elevation of serum heat-shock protein levels in amyotrophic lateral sclerosis." in: Neurological sciences : official journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology, Vol. 37, Issue 8, pp. 1277-81, (2016) (PubMed).

Tian, He, Fu, Lin, Tang, Huang, Guo, Sun: "High expression of heat shock protein 90 alpha and its significance in human acute leukemia cells." in: Gene, Vol. 542, Issue 2, pp. 122-8, (2014) (PubMed).

Target Details for HSP90

Target: HSP90 (Heat Shock Protein 90 (HSP90))

Alternative Name: HSP90 (HSP90 Products)

Synonyms: EL52 ELISA Kit, HSP86 ELISA Kit, HSP89A ELISA Kit, HSP90A ELISA Kit, HSP90N ELISA Kit, HSPC1 ELISA Kit, HSPCA ELISA Kit, HSPCAL1 ELISA Kit, HSPCAL4 ELISA Kit, HSPN ELISA Kit, Hsp89 ELISA Kit, Hsp90 ELISA Kit, LAP2 ELISA Kit, git10 ELISA Kit, swo1 ELISA Kit, HSP90 ELISA Kit, htpG ELISA Kit, SCBAC25F8.08 ELISA Kit, 23.m06066 ELISA Kit, 17.m07646 ELISA Kit, HSP90-1 ELISA Kit, 143198_at ELISA Kit, 83 ELISA Kit, 83K HSP ELISA Kit, DMHSP82 ELISA Kit, E(sev)3A ELISA Kit, E(sina)2 ELISA Kit, HSP82 ELISA Kit, HSP83 ELISA Kit, ORF1 ELISA Kit, Su(Raf)3A ELISA Kit, anon-EST:Liang-2.53 ELISA Kit, anon-WO0068693 ELISA Kit, anon-WO0140519.209 ELISA Kit, clone 2.53 ELISA Kit, en(lz)3C/4C ELISA Kit, hsp84 ELISA Kit, l(3)j5C2 ELISA Kit, ms(3)08445 ELISA Kit, stc ELISA Kit, DmelCG1242 ELISA Kit, CG1242 ELISA Kit, Hsp86 ELISA Kit, Hspca ELISA Kit, 86kDa ELISA Kit, 89kDa ELISA Kit, AL024080 ELISA Kit, AL024147 ELISA Kit, Hsp86-1 ELISA Kit, hsp4 ELISA Kit, hsp86 ELISA Kit, hsp89 ELISA Kit, hsp90 ELISA Kit, hsp90a ELISA Kit, hspc1 ELISA Kit, hspca ELISA Kit, hspn ELISA Kit, lap2 ELISA Kit, Hsp90alpha ELISA Kit, heat shock protein 90 alpha family class A member 1 ELISA Kit, heat shock protein Hsp90 ELISA Kit, Hsp90 chaperone ELISA Kit, Heat shock protein 90 ELISA Kit, heat shock protein 90 ELISA Kit, chaperone protein HtpG ELISA Kit, molecular chaperone HtpG ELISA Kit, Heat Shock Protein 90 ELISA Kit, uncharacterized LOC100384473 ELISA Kit, Heat shock protein 83 ELISA Kit, heat shock protein 90, alpha (cytosolic), class A member 1 ELISA Kit, heat shock protein 90 alpha family class B member 1 ELISA Kit, heat shock protein 90kDa alpha family class A member 1 L homeolog ELISA Kit, heat shock protein HSP 90-alpha ELISA Kit, HSP90AA1 ELISA Kit, hsp90 ELISA Kit, HSP90 ELISA Kit, daf-21 ELISA Kit, htpG ELISA Kit, SCO7516 ELISA Kit, MAP_RS10510 ELISA Kit, TP04_0646 ELISA Kit, TP01_0934 ELISA Kit, APH_RS03525 ELISA Kit, GbCGDNIH1_0315 ELISA Kit, MAV_2118 ELISA Kit, HSP90C ELISA Kit, BBOV_IV008400 ELISA Kit, BBOV_III007380 ELISA Kit, ACICU_00312 ELISA Kit, ECL_01244 ELISA Kit, YE105_C1172 ELISA Kit, pco153543(105) ELISA Kit, Hsp83 ELISA Kit, Hsp90aa1 ELISA Kit, HSP90AB1 ELISA Kit, hsp90aa1.1.L ELISA Kit, LOC108698781 ELISA Kit

Background: HSP90 is a highly conserved and essential stress protein that is expressed in all eukaryotic cells. From a functional perspective, HSP90 participates in the folding, assembly, maturation, and stabilization of specific proteins as an integral component of a chaperone complex. Despite its label of being a heat-shock protein, HSP90 is one of the most highly expressed proteins in unstressed cells (1-2 % of cytosolic protein). It carries out a number of housekeeping functions - including controlling the activity, turnover, and trafficking of a variety of proteins. Most of the HSP90- regulated proteins that have been discovered to date are involved in cell signaling. The number of proteins now known to interact with HSP90 is about 100. Target proteins include the kinases v-Src, Wee1, and c-Raf, transcriptional regulators such as p53 and steroid receptors, and the polymerases of the hepatitis B virus and telomerase(3). When bound to ATP, HSP90 interacts with co-chaperones Cdc37, p23, and an assortment of immunophilin-like proteins, forming a complex that stabilizes and protects target proteins from proteasomal degradation. In most cases, HSP90-interacting proteins have been shown to co-precipitate with HSP90 when carrying out immune-oadsorption studies, and to exist in cytosolic heterocomplexes with it. In a number of cases, variations in HSP90 expression or HSP90 mutation has been shown to degrade signaling function via the protein or to impair a specific function of the protein (such as steroid binding, kinase activity) in vivo. Ansamycin antibiotics, such as geldanamycin and radicicol, inhibit HSP90 function.

Pathways: M Phase, Regulation of Cell Size, Signaling Events mediated by VEGFR1 and VEGFR2, VEGFR1 Specific Signals