The Assay Genie Rabbit HP (Haptoglobin) ELISA Kit is a highly sensitive assay for the quantitative measurement of Rabbit HP in the following samples: Serum, Plasma, Cell Culture Supernatant, Cell or Tissue Lysate, Other Liquid Samples.
An HRP-conjugated antibody binds specifically to those recognizing HP. After washing, detection occurs via a colorimetric reaction where HRP oxidizes TMB, producing a blue signal that turns yellow upon addition of stop solution. The resulting optical density (OD) is measured at 450 nm using a microplate reader. To improve accuracy and account for potential optical interference, a reference wavelength of 650 nm may also be applied. The intensity of the yellow coloration directly correlates with the presence of HP in the tested sample.
This dual function kit includes validated Bradford Reagent to quantify total protein concentration for accurate sample normalization.
Product Name:
Rabbit HP (Haptoglobin) ELISA Kit
SKU:
AEFI01230
Reactivity:
Rabbit
Detection Method:
Capture
Sensitivity:
0.094 ng/mL
Range:
0.156-10 ng/mL
Sample Type:
Serum, Plasma, Cell Culture Supernatant, Cell or Tissue Lysate, Other Liquid Samples
Storage:
2-8°C for 12 months.
Linearity:
Sample
1:2
1:4
1:8
Serum (n = 5)
92-102%
82-91%
88-99%
EDTA Plasma (n = 5)
88-102%
86-95%
80-99%
Heparin Plasma (n = 5)
85-95%
83-95%
86-97%
Recovery:
Matrix
Recovery Range (%)
Average (%)
Serum (n = 5)
89-99
93
EDTA Plasma (n = 5)
86-103
94
Heparin Plasma (n = 5)
85-102
94
Note:The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Step
Procedure
1
Reagent & Plate Preparation: Equilibrate TMB substrate to room temperature for at least 30 minutes. Set standard, test sample and control (zero) wells on the pre-coated plate and record their positions. Each standard and sample is recommended to be measured in duplicate.
2
Standard, Sample & Control Loading: Aliquot 100 µl of standard solutions into the standard wells, 100 µl of properly diluted sample into test sample wells, and 100 µl of Sample Dilution Buffer into the control (blank) well.
3
First Incubation & Wash: Seal the plate and incubate at 37°C for 90 minutes. Aspirate the liquid and wash the plate 3 times with 350 µl Wash Buffer.
4
HRP-labelled Antibody & Wash: Add 100 µl of HRP-labelled working solution into each well, cover and incubate at 37°C for 30 minutes. Wash the plate 5 times with 350 µl Wash Buffer, allowing the buffer to stay in the wells for 1-2 minutes per wash.
5
Color Development: Add 90 µl of TMB substrate into each well and incubate at 37°C in the dark for 10-20 minutes.
6
Stop Reaction & Reading: Add 50 µl of Stop Solution and measure absorbance at 450 nm in a microplate reader immediately.
Sample Type
Protocol
Serum
Allow whole blood to coagulate at room temperature (2 h) or 2-8°C overnight. Centrifuge at 1000 × g for 20 min and collect the supernatant. Store or use immediately.
Plasma
Collect in anticoagulant tubes (EDTA, citrate, or heparin), mix gently, and centrifuge within 30 min at 1000 × g, 2-8°C for 15 min. Store or assay as needed.
Tissue Homogenate
Homogenize tissue in 50 mM Tris + 0.9% NaCl + 0.1% SDS (pH 7.3) with protease inhibitors (e.g. 1 mM PMSF). Avoid NP-40, Triton X-100 and DTT. Disrupt by ultrasonic homogenization or freeze-thaw cycles, centrifuge at 5000 × g for 5 min and collect the supernatant.
Cell Culture Supernatant
Centrifuge at 2500 rpm for 5 minutes at 2-8°C and collect the clarified cell culture supernatant.
Cell Lysate
Lyse cells in lysis buffer with protease inhibitor (e.g. 1 mM PMSF) on ice for 30-60 minutes, or by ultrasonic treatment. Centrifuge at 10,000 rpm for 10 min at 2-8°C and collect the supernatant.
Other Sample Types
For more information about how to process other sample types, (e.g., body fluids, breast milk & more), please contact our Tech Support Team at techsupport@assaygenie.com.
Component
Quantity
Storage
48T
96T
ELISA Microplate (Dismountable)
8×6
8×12
Place the test strips into a sealed foil bag with the desiccant. Store for 1 month at 2-8°C; Store for 12 months at -20°C.
Lyophilized Standard
1 vial
2 vial
Place the standards into a sealed foil bag with the desiccant. Store for 1 month at 2-8°C; Store for 12 months at -20°C.
