The Human F-Hb Free Hemoglobin ELISA Kit is a reliable and accurate tool for the detection of free hemoglobin levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and reproducible results for a variety of research applications. Free hemoglobin is a key indicator of hemolysis and can be a critical biomarker in conditions such as hemolytic anemia, blood transfusion reactions, and various other hematological disorders.
The ability to accurately measure free hemoglobin levels can provide valuable insights into disease progression, treatment response, and overall patient health. With the Human F-Hb Free Hemoglobin ELISA Kit, researchers can confidently study and monitor free hemoglobin levels in a convenient and efficient manner, advancing their understanding of hemolytic disorders and improving patient care.
Product Name:
Human f-Hb/Free Haemoglobin ELISA Kit
SKU:
HUFI02466
Reactivity:
Human
Assay Type:
Sandwich ELISA, Double Antibody
Sensitivity:
0.188 ng/mL
Range:
0.313-20 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)
85-101%
87-100%
87-99%
EDTA Plasma (n = 5)
88-104%
88-97%
80-93%
Heparin Plasma (n = 5)
86-103%
82-101%
84-94%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 5)
85-102
96
EDTA Plasma (n = 5)
87-102
94
Heparin Plasma (n = 5)
86-104
95
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 reagents and TMB substrate to room temperature. Set standard, test sample and control (zero) wells on the pre-coated plate and record their positions.
2
Primary Incubation: Prepare standards, samples, blanks and load into designated wells. Incubate plate at 37°C for 90 minutes to allow antigen binding.
3
Detection Antibody Binding: Add biotin-labeled detection antibody and incubate at 37°C for 60 minutes.
4
HRP-Streptavidin Binding: Add HRP-Streptavidin (SABC) and incubate at 37°C for 30 minutes.
5
Color Development: Add TMB substrate and incubate in the dark for 10–20 minutes.
6
Stop Reaction & Reading: Add stop solution and measure absorbance at 450 nm immediately.
Sample Type
Protocol
Serum
Allow blood to clot, centrifuge at 1000 × g for 20 minutes, collect supernatant supernatant and store appropriately.
Plasma
Collect using anticoagulant tubes, centrifuge at 1000 × g for 15 minutes at 2–8°C and collect plasma.
Tissue Homogenate
Homogenize tissue in PBS with protease inhibitors, centrifuge and collect supernatant.
Cell Culture Supernatant
Centrifuge at 2500 rpm for 5 minutes and collect clarified supernatant.
Cell Lysate
Lyse cells using lysis buffer with protease inhibitors, centrifuge and collect protein 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.
