Human ErbB2/Her2 Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
DHER20
Control Products Available
Human ErbB2/Her2 Standard Curve
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Human ErbB2/Her2 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Urine (50 uL), Human Milk (50 uL)
Sensitivity
14.8 pg/mL
Assay Range
78.1 - 5,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine, Human Milk)
Specificity
Natural and recombinant human ErbB2.
Cross-reactivity
Cross-reactivity observed with 1 or more available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human ErbB2/Her2 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human ErbB2 in cell culture supernates, cell lysates, serum, plasma, urine, and human milk. It contains NS0-expressed recombinant human ErbB2 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human ErbB2 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human ErbB2.

Precision

Intra-Assay Precision (Precision within an assay) Intra-assay Precision (Precision within an assay)<br>Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Inter-assay Precision (Precision between assays)<br>Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 471 1379 2681 489 1419 2667
Standard Deviation 6.71 16.3 31.1 18.7 56.4 93.5
CV% 1.4 1.2 1.2 3.8 4 3.5

Recovery

The recovery of human ErbB2 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 103 90-120
EDTA Plasma (n=4) 99 89-118
Heparin Plasma (n=4) 104 92-119
Serum (n=4) 99 91-115
Urine (n=4) 101 94-106

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human ErbB2 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human ErbB2/Her2 ELISA Linearity

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: ErbB2/Her2

ErbB2, also called Neu and Her2, is a transmembrane glycoprotein in the ErbB family of tyrosine kinase receptors for EGF superfamily growth factors. ErbB2 is widely expressed in epithelial cells and over-expressed in a large number of breast carcinomas. ErbB2 has no identified ligands but heterodimerizes with ErbB1/EGF R, ErbB3, or ErbB4 to form higher affinity signaling complexes. The protease ADAM10 releases a 110 kDa soluble fragment of ErbB2 from the cell surface. ErbB2 plays roles in development, cancer, communication at the neuromuscular junction, and regulation of cell growth and differentiation. The ErbB2/ErbB3 heterodimer is expressed in the majority of breast, skin, ovary and gastrointestinal tumors and transduces a highly mitogenic signal in response to neuregulin 1 (NRG1; heuregulin 1) or NRG2. ErbB3, ErbB2 and neuregulin are all required for formation of the sympathetic nervous system.

Long Name:
Receptor Tyrosine Protein Kinase ErbB2
Entrez Gene IDs:
2064 (Human); 13866 (Mouse); 24337 (Rat)
Alternate Names:
CD340 antigen; CD340; c-erb B2/neu protein; EC 2.7.10; EGFR2; ErbB2; HER2; HER-2; HER2EC 2.7.10.1; herstatin; Metastatic lymph node gene 19 protein; MLN 19; MLN19; Neu Oncogene; NEUHER-2/neu; neuroblastoma/glioblastoma derived oncogene homolog; NGL; NGLTKR1; p185erbB2; Proto-oncogene c-ErbB-2; Proto-oncogene Neu; receptor tyrosine-protein kinase erbB-2; TKR1; Tyrosine kinase-type cell surface receptor HER2; v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 2(neuro/glioblastoma derived oncogene homolog); v-erb-b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastomaderived oncogene homolog (avian)
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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