Teacher Notes

ABO/Rh Simulated Blood Typing

Student Laboratory Kit

Materials Included In Kit

Blood sample, Person W, 30 mL
Blood sample, Person X, 30 mL
Blood sample, Person Y, 30 mL
Blood sample, Person Z, 30 mL
Simulated Anti-A Sera, 30 mL
Simulated Anti-B Sera, 30 mL
Simulated Anti-Rh Sera, 30 mL
Blood typing slide wells, 50
Toothpicks, 100

Additional Materials Required

Marking pens

Safety Precautions

The solutions in this simulation are considered non-hazardous but normal safe laboratory procedures should be followed. Wear chemical splash goggles, chemical-resistant gloves and a chemical-resistant apron. Please review current Safety Data Sheets for additional safety, handling and disposal information. Remind students to wash their hands thoroughly with soap and water before leaving the laboratory.


Please consult your current Flinn Scientific Catalog/Reference Manual for general guidelines and specific procedures, and review all federal, state and local regulations that may apply, before proceeding. Dispose of all products down the drain according to Flinn Suggested Disposal Method #26b.

Teacher Tips

  • The theory of blood chemistry and blood type genetics as outlined in the background should be studied and discussed prior to doing this lab activity.

  • This lab does not use any real blood components and therefore, poses no greater risk than the handling of the small quantities of chemicals involved in the tests.
  • Similar to real blood, the simulated blood does contain solids. Do not shake the bottles as the solids will clog the dropping tip. Use a dissecting needle to clear any clogged tips. In this case the solids are due to a reaction between the red coloring and the calcium in two of the simulated blood samples.
  • Anti-A and Rh sera have sodium sulfate as the ion that precipitates with the calcium in the simulated blood samples for the positive test. This precipitation requires between two to four minutes to produce a clearly visible positive test.

Correlation to Next Generation Science Standards (NGSS)

Science & Engineering Practices

Asking questions and defining problems
Developing and using models
Planning and carrying out investigations
Analyzing and interpreting data
Engaging in argument from evidence
Obtaining, evaluation, and communicating information

Disciplinary Core Ideas

MS-LS1.A: Structure and Function
MS-PS1.B: Chemical Reactions
HS-PS1.B: Chemical Reactions
HS-LS1.A: Structure and Function
HS-LS3.A: Inheritance of Traits

Crosscutting Concepts

Cause and effect
Systems and system models
Structure and function

Performance Expectations

MS-LS1-1. Conduct an investigation to provide evidence that living things are made of cells; either one cell or many different numbers and types of cells
MS-LS1-2. Develop and use a model to describe the function of a cell as a whole and ways parts of cells contribute to the function.
MS-PS1-2. Analyze and interpret data on the properties of substances before and after the substances interact to determine if a chemical reaction has occurred.
HS-PS1-2. Construct and revise an explanation for the outcome of a simple chemical reaction based on the outermost electron states of atoms, trends in the periodic table, and knowledge of the patterns of chemical properties.
HS-LS1-2. Develop and use a model to illustrate the hierarchical organization of interacting systems that provide specific functions within multicellular organisms.
HS-LS1-1. Construct an explanation based on evidence for how the structure of DNA determines the structure of proteins, which carry out the essential functions of life through systems of specialized cells.
HS-LS3-1. Ask questions to clarify relationships about the role of DNA and chromosomes in coding the instructions for characteristic traits passed from parents to offspring.

Sample Data


Answers to Questions

  1. What is Person X’s blood type? What antigens are present on the surface of the red blood cells in Person X?

    Person X is type O. Person X, therefore, has no antigens on the surface of the red blood cells.

  2. What is Person Y’s blood type? What antibodies are present in Person Y’s plasma?

    Person Y has Type A blood. Person Y has b-antibodies in their plasma.

  3. Person Z needs a transfusion. What blood types might Person Z safely receive? Explain.

    Person Z has blood type B. Person Z can safely receive blood type B blood since it matches. Person Z could also receive blood type O (the universal donor blood) if transfused slowly.

  4. Could a man with type AB blood be the father of a child with type O blood? Explain.

    No, the man with type AB blood could not have a child with type O blood. The man would produce the alleles IA and IB and give one or the other to any offspring. Type O blood has the ii genotype and therefore, could not be the offspring of a type AB.

  5. Could a child with type B blood with a mother of type A blood have a father with type A blood? Explain.

    No, the child would have to get the IB allele from one of the parents. Type A parents only produce IA or i alleles and therefore, produce no IB alleles.

Student Pages

ABO/Rh Simulated Blood Typing


A blood transfusion with blood of a mismatched blood type usually has serious consequences for the recipient of the blood. Today, complete blood analysis is done with sophisticated, costly equipment before transfusions are done. The basic principles of blood typing will be illustrated in this activity using simulated ABO and Rh blood typing sera and simulated bloods.


  • Antigens

  • Multiple alleles
  • Rh factor
  • Antibodies
  • Codominance



Early attempts to transfer blood from one person to another produced varied results. Sometimes it seemed to help the recipient and other times it produced very serious consequences. Eventually, it was discovered that each individual has a unique combination of substances in his or her blood. Some of these substances may be compatible with another person’s blood and some may not be compatible. These findings led to the discovery and development of procedures to type an individuals’ blood. It is now known that safe transfusions of blood depend upon properly matching the blood types of the donors and the recipients.

Genetics of Blood Types

ABO blood type is determined by the presence or absence of specific proteins on an individual’s red blood cells. A basic genetic principle is that an individual’s inherited genes determines which proteins are produced in the individual’s body. In the ABO blood typing system (just one of many blood factors) the blood proteins (antigens) are called the A and B proteins. The presence or absence of the A and B proteins on the red blood cells determines the individual’s blood type in the ABO typing system. Individuals whose red blood cells contain protein A and lack protein B have type A blood. Those with protein B and lack protein A are called type B. Individuals with both protein A and protein B are called type AB and individuals with neither of the proteins is called type O.

ABO blood type is a genetic example of multiple alleles. There are three alleles in the gene pool for ABO blood type (i.e., IA, IB, i). IA codes for protein A, IB codes for protein B and i codes for neither protein A nor protein B. Within this multiple allele pool the gene interactions illustrate both simple dominance as well as codominance. (Remember each individual has only two alleles for each trait even if there are multiple alleles in the gene pool.) When the IAi allele combination occurs, the individual is blood type A. When the IAIB combination occurs, the IA and IB alleles are codominant and the individual is blood type AB. The following chart illustrates the allele combinations, resulting blood type, proteins on the red blood cells, and antibodies in the blood for the four blood types in the ABO system.


Blood Transfusions

Blood groups are critically important with respect to transfusions. If someone with type A is given a transfusion of type B blood the two bloods will interact, clump and clog arteries which will have serious consequences to the individual. The clumping reaction is caused by the interaction of the proteins on the red blood cells and the antibodies present in the blood plasma. Antibodies are produced by the body in reaction to foreign proteins and are important in protecting the body against disease. Antibodies cannot distinguish a disease protein from protein on red blood cells. Individuals do not produce antibodies for pro-teins of their own red blood cells, but do produce antibodies for foreign proteins. Thus, a person with type A blood (A protein on surface of red blood cells) does not produce a antibodies. This person does produce b antibodies. If given the transfusion of type B blood, the antigens and antibodies of the mismatched blood will react and clump (a natural defense mechanism for foreign proteins). The illustrations below, in a very oversimplified way, illustrate the makeup of each of the four blood types.


Using the same illustration scheme, a transfusion of type B blood into an individual with type A blood might be illustrated as follows:


Similarly, a person with type B blood must not be given a transfusion with type A blood.

Because type AB blood lacks both a and b antibodies, it would appear that an AB person could receive a transfusion of blood from any other type. For this reason, type AB persons are sometimes called universal recipients. It should be noted, however, that type A (b), type B (a) and type O (a and b) blood still contain antibodies (either a or b) that could cause clumping of type AB cells. Consequently, even for AB individuals, it is always best to use donor blood of exactly the same type as the recipient blood. If the matching type is not available and type A, B or O is used, it should be transfused very slowly so that the donor blood is well diluted by the recipient’s larger blood volume.

Similarly, because type O blood lacks antigens A and B, it would seem that this blood type could be transfused into persons with blood of any other type. For this reason, persons with type O blood are often referred to as universal donors. Type O blood, however, does contain both anti-a and anti-b antibodies, and thus, if it is transfused into a person of a different blood type it should be done slowly to minimize large clumping reactions.

The bottom line for transfusion is that blood types should be matched for transfusions.

Blood Typing

ABO blood typing is based upon the clumping phenomena of bloods of mixed types. Blood sera antibodies can be isolated from other components of the blood and then used as blood typing sera. Antibodies-b (called Anti-a sera), for example, would clump red blood cells containing A-antigens (type A). Anti-b sera would clump type B blood. Clumping will occur in both sera with type AB blood and in neither sera with type O blood.

In the ABO blood typing procedure, drops of blood are first secured following sterile procedures. A drop of blood is placed in a drop of anti-a sera and another drop is placed in a drop of anti-b sera. The drops are then observed for clumping. The pattern of clumping or non-clumping is interpreted and the blood type determined. The following patterns occur for the various blood types:


There are many other blood typing systems in addition to the ABO classification system. One commonly used system is the Rh factor.

The Rh blood group has several antigen factors on the surface of the red blood cells. If any of the antigens are present on the RBC surface, clumping can occur and the individual is said to be Rh positive (Rh+). Conversely, if the red cells lack Rh antigens, the blood is said to be Rh negative (Rh).

Just like the ABO system the Rh factors are inherited. The genetics follow a simple dominant/recessive inheritance with Rh+ being dominant. Unlike anti-a and anti-b, antibodies for Rh (anti-Rh) do not appear spontaneously. Instead, they form only in Rh persons in response to special stimulation.

If an Rh person receives a transfusion of Rh+ blood, the recipient’s antibody producing cells are stimulated by the presence of the foreign Rh antigen and will begin producing anti-Rh antibodies. Generally, no serious consequences result from this initial transfusion. But if the Rh person (who is now sensitized to Rh+ blood—has antibodies) received another transfusion of Rh+ blood at a later time, the donor’s red cells are likely to clump.

A related condition may occur when an Rh woman is pregnant with an Rh+ fetus for the first time. Such a pregnancy may be uneventful if the fetus’ blood and mother’s blood do not mix during birth. If, however, during birth or miscarriage, some of the infant’s Rh+ blood cells get into the mother’s Rh blood, it might sensitize her blood and begin the production of anti-Rh antibodies.

If a mother who has already developed anti-Rh antibodies becomes pregnant with a second Rh+ fetus, the anti-Rh antibodies can pass through the placental membranes and react with the Rh+ fetal red cells, causing them to clump. The fetus often develops a condition known as erythroblastosis fetalis, which is likely to be very serious for the fetus.


(for each lab group)
Blood sample, Person W, 9–12 drops
Blood sample, Person X, 9–12 drops
Blood sample, Person Y, 9–12 drops
Blood sample, Person Z, 9–12 drops
Simulated Anti-A Sera, 12–16 drops
Simulated Anti-B Sera, 12–16 drops
Simulated Anti-RH Sera, 12–16 drops
Blood typing slide wells, 4
Marking pen
Toothpicks, 12

Safety Precautions

Wear chemical splash goggles, chemical-resistant gloves and a chemical-resistant apron. Wash hands thoroughly with soap and water before leaving the laboratory.


{10198_Procedure_Figure_3_Blood typing slide well}
  1. Label four blood typing slide wells—“W,” “X,” “Y” and “Z” using a marking pen (see Figure 3).
  2. Place 3–4 drops of Person W blood in each of the three wells in the slide marked “W.” Similarly place 3–4 drops of “X” blood in each well on slide “X,” “Y” blood on slide “Y” and “Z” blood on slide “Z.” Note: Do not shake the bottles.
  3. Add 3–4 drops of Anti-A Sera to each slide in the Anti-A well. Similarly, add Anti-B Sera to each Anti-B well and Anti-Rh to each Anti-Rh well.
  4. Stir the mixtures in all 12 wells being careful not to scratch the plastic. Use a different clean toothpick for each well. Use only one toothpick per well to avoid cross contamination. Mix each solution thoroughly and let the slides sit for at least three minutes.
  5. Observe each well against a white background (paper) and record the results on the ABO/Rh Blood Typing Worksheet.
  6. Study the results and answer the questions on the ABO/Rh Blood Typing Worksheet.
  7. Dispose of all materials as directed by your instructor.

Student Worksheet PDF


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