QuantiChrom™ Hemoglobin Assay Kit
Datasheet PDF  MSDS/Bulk Order


Application
For quantitative determination of hemoglobin and evaluation of drug effects on hemoglobin metabolism.

Key Features
Sensitive and accurate. Linear detection range 0.9 - 200 mg /dL hemoglobin in 96-well plate assay.

Simple and high-throughput. The "mix-and-read" procedure involves addition of a single working reagent and reading the optical density. Can be readily automated as a high-throughput assay in 96-well plates for thousands of samples per day.

Safety. Reagents are non-toxic.

Versatility. Assays can be executed in 96-well plate or cuvet.

Method
OD400nm

Samples
Blood, serum, plasma, urine etc

Species
All

Procedure
5 min

Size
250 tests

Detection Limit
0.9 mg/dL

Shelf Life
12 months

More Details
HEMOGLOBIN (Hb) is made of four globin chains each carrying a heme group. It is carried by red blood cells and transports oxygen from the lungs to the peripheral tissues to maintain the viability of cells. Quantitation of blood hemoglobin has been a key diagnostic parameter for various diseases such as anemia, polycythemia and dehydration. Simple, direct and automation-ready procedures for measuring hemoglobin concentration are becoming popular in Research and Drug Discovery. BioAssay Systems’ QuantiChrom™ hemoglobin assay kit is based on an improved Triton/NaOH method, in which the hemoglobin is converted into a uniform colored end product. The intensity of color, measured at 400 nm, is directly proportional to hemoglobin concentration in the sample. The optimized formulation exhibits high sensitivity and is ideal for measuring hemolysis in low hemoglobin samples (e.g. serum and plasma).

1. The procedure mentions that the Standard is to be diluted with water and NO reagent addition. Is the standard already converted cyanohemiglobin? How does this standard reflect OD differences after different incubation times? Wouldn’t you want to create a standard curve for extrapolation of your sample values?



Our assay does not use the cyanohemoglobin method. The "Standard" is not hemoglobin itself as hemoglobin is unstable. We are using a dye at a concentration that gives the same OD reading as the freshly prepared hemoglobin at 100 mg/dL. Therefore when you apply the "Standard", there is no need to use the reagent.

The changes in OD with incubation time is small if any, but we recommend reading the OD values at 5 min after adding the reagent. The standard curve shown in the protocol was read using a fresh pure hemoglobin preparation and shows that the assay is linear up to 200 mg/dL hemoglobin. Thus there is no need to construct a full standard curve. We recommend using the blank and 100 mg/dL Calibrator to calculate the sample hemoglobin concentration. As long as hemoglobin values are below 200 mg/dL, there is no concern about non-linearity. If it is > 200 mg/dL, we recommend diluting the sample in water and repeat the assay, multiply the value by the dilution factor.

2. Why can you determine the concentration of hemoglobin in plasma using the kit? I was wondering why the color of plasma (yellow) would not interfere with the measurement of the absorbance at 400 nm. Do you have any data to show that plasma does not interfere with the measurement?



The yellow color of serum is caused mostly by hemoglobin and bilirubin and our assay does not distinguish between the two. However, normal bilirubin levels will add only a relatively minor positive bias which will not elevate the reading above normal hemoglobin levels in serum (< 10 mg/dL). This kit is suitable to test for elevated hemoglobin levels in serum due to hemolysis. If you need to accurately measure the exact hemoglobin concentration in normal serum samples, you probably should take into account the background added by bilirubin. For such applications other assay kits may be more suitable.

For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions
1. Thaker PH et al (2006). Chronic stress promotes tumor growth and angiogenesis in a mouse model of ovarian carcinoma. Nat Med. 12(8):939-44. Assay: Hemoglobin in Mouse tumor tissue (Pubmed).

2. Liu D et al (2010). Puma is required for p53-induced depletion of adult stem cells. Nat Cell Biol. 12(10):993-8. Assay: Hemoglobin in Mouse tissue (Pubmed).

3. Burne-Taney MJ et al (2006). Decreased capacity of immune cells to cause tissue injury mediates kidney ischemic preconditioning. J Immunol. 176(11):7015-20. Assay: Hemoglobin in Mouse spleen cell (Pubmed).

4. Kasiappan R et al (2009). Loss of p53 and MCT-1 overexpression synergistically promote chromosome instability and tumorigenicity. Mol Cancer Res. 7(4):536-48. Assay: Hemoglobin in Mouse tumor tissue (Pubmed).

5. Liu W,et al (2009). Normobaric hyperoxia reduces the neurovascular complications associated with delayed tissue plasminogen activator treatment in a rat model of focal cerebral ischemia. Stroke 40(7):2526-31. Assay: Hemoglobin in Rat tissue (Pubmed).

6. Weis SM et al (2008). Compensatory role for Pyk2 during angiogenesis in adult mice lacking endothelial cell FAK. J Cell Biol. 181(1):43-50. Assay: Hemoglobin in Mouse tumor tissue (Pubmed).

7. He Y et al (2009). Effects of cerebral ischemia on neuronal hemoglobin. J Cereb Blood Flow Metab. 29(3):596-605. Assay: Hemoglobin in Rat brain tissue (Pubmed).

8. Fridén M et al (2010). Improved measurement of drug exposure in the brain using drug-specific correction for residual blood. J Cereb Blood Flow Metab. 30(1):150-61. Assay: Hemoglobin in Rat brain tissue (Pubmed).

9. Meyer C et al (2010). Hemodialysis-induced release of hemoglobin limits nitric oxide bioavailability and impairs vascular function. J Am Coll Cardiol. 55(5):454-9. Assay: Hemoglobin in Human plasma (Pubmed).

10. Singleton PA et al (2010). Methylnaltrexone potentiates the anti-angiogenic effects of mTOR inhibitors. J Angiogenes Res. 2(1):5. Assay: Hemoglobin in Mouse tumor tissue (Pubmed).

11. French CJ et al (2010). Vascular rhexis: loss of integrity of coronary vasculature in mice subjected to myocardial infarction. Exp Biol Med (Maywood) 235(8):966-73. Assay: Hemoglobin in Mouse tissue (Pubmed).

To find more recent publications, please click here.
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QuantiChrom™ Hemoglobin Assay Kit
Catalog No: DIHB-250
Price: $289    Qty:
For orders of 10 or more kits, please contact us at 1-510-782-9988 x 1 for best pricing.

Shipping: RT
Shipment: Same Day for order by 2pm PST
Delivery: 1-2 days (US), 3-6 days (Intl);
Storage: 4°C



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