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Mammalian Protamines

$100.00$1,700.00

Native and synthetic forms of the mammalian protamine P1 and P2 proteins are available

  • Native protamine molecules:
    • Protamine P1 as isolated from sperm (bull, boar, ram)
    • Protamine P1 isolated from sperm and then purified by HPLC (bull and hamster)
    • Protamine P1P2 Mix as isolated from sperm (hamster, stallion, and human)
  • Synthetic HPLC purified human, stallion and mouse protamines

If you are interested in protamines isolated from the sperm of a species other than those listed here, please contact us – we may be able to help.

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Description

Our products are sold for research use only and are not intended for diagnostic or therapeutic use in humans.

Protamines are small, arginine-rich, nuclear proteins that replace histones and transition proteins late in spermiogenesis. They play an important role in spermatid genome inactivation and sperm chromatin condensation and stabilization. Protamine binding to DNA may contribute to the forces that drive nuclear compaction and contribute to head shaping. DNA bound to protamines is much more densely packed than DNA packaged by histones, and the genes associated with protamine can only be expressed when the protamines are removed after fertilization.

Native Mammalian Protamines

Bull (Bos taurus) P1 (MW 6627) is a native protamine that has been isolated from mature sperm. Two forms are available – the native protamine as isolated from sperm and the isolate further purified by HPLC. Bull P1 protamine is recognized by and binds to the Hup1N antibody. It does not bind to the Hup2B or the Hup1M antibodies.

Boar (Sus scrofa) P1 and Ram Ram (Ovis aries) P1 are native protamines that have been isolated from mature sperm. These native P1 molecules are recognized by and bind to the Hup1N antibody. They do not bind to the Hup2B or the Hup1M antibodies.

Hamster (Mesocricetus auratus) P1 is a native protamine that has been isolated from mature sperm and then purified by HPLC. Hamster P1 protamine is recognized by and binds to the Hup1N antibody. It does not bind to the Hup2B or the Hup1M antibodies.

Hamster (Mesocricetus auratus) P1P2 mix has been isolated from mature sperm. These native protamines are present in hamster sperm as a mixture (P1: 67% and P2 33%). The hamster P1 and P2 protamines are recognized by the Hup1N and Hup2B antibodies, respectively. Neither protamine binds to the Hup1M antibody.

Stallion (Equus caballus) P1P2 mix has been isolated from mature sperm. These native protamines are present in stallion sperm as a mixture (P1: 86% and P2 14%). The stallion P1 protamine is recognized by the Hup1N antibody. Stallion P2 does not bind to Hup2B. Neither protamine binds to the Hup1M antibody.

Human (Homo sapiens) P1P2 mix has been isolated from mature sperm. These native protamines are present in human sperm as a mixture (P1: 50% and P2: 50%). The human P1 protamine is recognized by Hup1N and Hup1M. The human P2 protamine is recognized by Hup2B.

Synthetic Mammalian Protamines

Human (Homo sapiens) synthetic P1 has been chemically synthesized, HPLC purified and analyzed by mass spectroscopy (95.52% purity, MW 6692.00). The synthetic human P1 is recognized by the Hup1N and Hup1M antibodies. It does not bind to the Hup 2B antibody.

Human (Homo sapiens) synthetic P2 has been chemically synthesized, HPLC purified and analyzed by mass spectroscopy (95.21% purity, MW 7655.05). The synthetic human P2 is recognized by the Hup2B antibody. It does not bind to the Hup1N and Hup1M antibodies.

Mouse (Mus musculus) synthetic P1 has been chemically synthesized, HPLC purified and analyzed by mass spectroscopy (96.7% purity, MW 6827.31). The synthetic mouse P1 is recognized by the Hup1N antibody. It does not bind to the Hup 2B and Hup1M antibodies.

Mouse (Mus musculus) synthetic P2 has been chemically synthesized, HPLC purified and analyzed by mass spectroscopy (95.2% purity, MW 8594.17). The synthetic human P2 is recognized by the Hup2B antibody. It does not bind to the Hup1N and Hup1M antibodies.

Stallion (Equus caballus) synthetic P1 has been chemically synthesized, HPLC purified and analyzed by mass spectroscopy (95.94% purity, MW 6,599.30).  It can be recognized by Hup1N.

Stallion (Equus caballus) synthetic P2 has been chemically synthesized, HPLC purified and analyzed by mass spectroscopy (77.84% purity, MW 8,040.56). It can not be recognized by any of the Hup antibodies.  We will soon be offering new antibodies on our website, including one that can recognize stallion P2.  Stay tuned.

 

Concentration

The protamines are dissolved in 100 mM 2-mercaptoethanol and are provided at a concentration of 1.0 mg/ml.

If you need to produce a more concentrated protamine stock solution, one of the following three approaches should work well.

  1. Transfer the protamine stock solution supplied (1.0 mg/ml) to a small Pyrex or equivalent tube and freeze on dry ice. Lyophilize to remove the solvent. The protein can then be dissolved in distilled water containing 10-100 mM 2-mercaptoethanol (MSH) or 10 mM HCL to the desired concentration.
  2. Transfer the protamine stock solution supplied (1.0 mg/ml) into a SpeedVac microcentrifuge tube and centrifuge the sample under vacuum in a SpeedVac centrifuge until the desired volume and concentration is reached.
  3. Mix the protamine stock solution with an equal volume of 40% trichloroacetic acid (TCA) to produce a final TCA concentration of 20% and allow the protein to precipitate on ice for 1 hr. Centrifuge to sediment the precipitated protamine, carefully remove the supernatant, and add 2-3 ml acidified acetone (2 drops 1N HCL added to 10 ml acetone) to rinse the protamine pellet, centrifuge again, carefully remove the supernatant, and repeat the acidified acetone wash twice more. After removing the acetone supernatant the final time, let the protein pellet air-dry. It is important to note that if small amounts of protamine are being precipitated in this manner, the resulting precipitate can be very difficult to see. It may appear as a barely visible white coating or a clear “glass-like” film on the bottom of the tube. The protein can then be dissolved in distilled water containing 10-100 mM 2-mercaptoethanol (MSH) or 10 mM HCL to the desired concentration.

Storage

These proteins can be stored as provided for up to six months at 4-6 °C. If they are to be maintained for longer periods of time, store at -20°C. Protamines stored in the dried state form intramolecular disulfides over time and can become insoluble. The formation of these intermolecular disulfides can be minimized by storing the dried proteins under an inert gas, such as nitrogen or argon. If  disulfides form, they can be reduced by adding concentrated 2-mercaptoethanol to the dry protein, allowing reduction of the disulfides to proceed for 30 min, and then adding water to adjust the protein and MSH concentration.

 

Additional information

Weight 0.1 lbs
Dimensions 0.1 x 0.1 x 0.1 in
Species/Protamine Type

Bull native P1, Bull native P1 HPLC, Boar native P1, Ram native P1, Hamster native P1 HPLC, Hamster native P1P2 mix, Stallion native P1P2 mix, Human native P1P2 mix, Human synthetic P1, Human synthetic P2, Mouse synthetic P1, Mouse synthetic P2, Stallion synthetic P1, Stallion synthetic P2

Quantity (micrograms)

100ug, 200ug, 500ug

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