- EG Cash
- 84,001
Can't take credit for this, as I've never tried this. But, the need for this recipe has presented itself, so here it is.
Description:
Recombinant Human Long-R3-IGF-I produced in E. coli is a single, non-glycosylated, polypeptide chain containing 83 amino acids and having a molecular mass of 9111 Dalton.
0.6% Acetic Acid Overview
Acetic Acid (AA) will be used to reconstitute (turn your lyophilized IGF-1 into a liquid form) your IGF-1. The standard is to use 0.6% AA. This concentration is typically not available for you to purchase. You can make your own 0.6% AA and I will show you how below (many have used this method successfully).
Making 0.6% Acetic Acid
You will have to purchase a few items upfront. Here is a ***8220;grocery list***8221; of items you will need. I have provided check boxes for you to check off once you have purchased these items.
Items Needed:
***8226; Distilled white vinegar (grocery store)
***8226; Distilled water (grocery store)
***8226; 0.2-0.22um sterile Whatman syringe filter
***8226; 10mL syringe with a luer lock tip
***8226; ~20-22 gauge needles (just the needles)
***8226; Sterile glass vial (10-20mL)
***8226; Alcohol prep pads ***8211; sterile kind (70% isopropyl alcohol)
Quick Guide:
1. Swab the top of your sterile vial with alcohol prep pad (70% isopropyl alcohol)
2. Mix 7.5mL distilled water with 1.0mL vinegar
3. Add Whatman syringe filter
4. Add sterile ~20ga. needle to end of Whatman filter
5. Inject the 8.5mL of solution into the sterile vial
6. You now have sterile 0.6% acetic acid
Detailed Directions:
1. Wash you hands thoroughly
2. Optional: wear alcohol treated exam gloves (rub your gloved hands together with 70% isopropyl alcohol on them until dry)
3. Using a sterile alcohol prep pad, swab the top of your sterile glass vial (into which the acetic acid solution will be held in)
4. Using the 10mL syringe with a ~20ga. needle on the end, draw up 7.5mL distilled water
5. Using the same syringe, now draw up 1mL vinegar
6. Remove needle from the syringe and discard
7. Attach 0.2-0.22um Whatman sterile syringe filter (do not touch the free end that will have a needle on it)
8. Put a new, sterile needle (~20 gauge) onto the free end of the Whatman filter (do not touch needle)
a. Do not use the same needle on the Whatman that was used to originally draw up the unsterile vinegar and distilled water.
9. Put a ~20 gauge sterile needle into the top of your sterile glass vial to act as a vent
10. Inject the acetic acid solution into the vial
11. You are now done and should have sterile 0.6% acetic acid
Notes:
1. These items MUST be sterile: 20-22ga. Needles, whatman filter, glass vial
2. Whatman filter: These small, sterile filters are used to filter the acetic acid solution so it is sterile. It does not matter that the liquid in your syringe (distilled water & vinegar) is not sterile, nor does it matter that the syringe itself is not sterile. Once the liquid goes through the filter it is STERILE. Thus, everything after the filter must be sterile!
3. You will most likely use 1mL (milliliter) of 0.6% AA to reconstitute your IGF-1. Thus, you should make at least 1.5mL. In reality, it***8217;s just as easy to make 8.5mL as I have stated in the above directions. You will have plenty for use later then.
4. Do NOT reuse the Whatman filter nor any needles! Discard immediately.
Reconstituting IGF-1lr3
Reconstitution is simply the addition of the 0.6% AA to your lyophilized IGF-1.
Assumption: 1mg/mL IGF-1/AA (1mg IGF-1 will be combined with 1mL AA; 1mg IGF-1 is the same as 1,000mcg)
1. Swab the top of your IGF-1 vial with a sterile alcohol prep pad
2. Swab the top of your 0.6% AA vial with a sterile alcohol prep pad
3. Using either multiple insulin syringe volumes (example: 2 x 0.5cc) or a single large syringe, obtain 1.0mL of 0.6% AA.
4. In the IGF-1 vial, insert a sterile ~20 ga. needle to act as a vent
5. Inject the 1.0mL of AA very slowly and dribble it down the side of the vial.
a. Be very careful with this peptide as it is very delicate!
6. Remove the needle & syringe and discard
7. Gently swirl the vial or roll between your hands.
a. Again, be very gentle here
8. You now have 1mg/mL of IGF-1
a. This is the same as: 1,000mcg/mL
Notes:
1. If you added 2mL of AA, it would be a 0.5mg/mL
Description:
Recombinant Human Long-R3-IGF-I produced in E. coli is a single, non-glycosylated, polypeptide chain containing 83 amino acids and having a molecular mass of 9111 Dalton.
0.6% Acetic Acid Overview
Acetic Acid (AA) will be used to reconstitute (turn your lyophilized IGF-1 into a liquid form) your IGF-1. The standard is to use 0.6% AA. This concentration is typically not available for you to purchase. You can make your own 0.6% AA and I will show you how below (many have used this method successfully).
Making 0.6% Acetic Acid
You will have to purchase a few items upfront. Here is a ***8220;grocery list***8221; of items you will need. I have provided check boxes for you to check off once you have purchased these items.
Items Needed:
***8226; Distilled white vinegar (grocery store)
***8226; Distilled water (grocery store)
***8226; 0.2-0.22um sterile Whatman syringe filter
***8226; 10mL syringe with a luer lock tip
***8226; ~20-22 gauge needles (just the needles)
***8226; Sterile glass vial (10-20mL)
***8226; Alcohol prep pads ***8211; sterile kind (70% isopropyl alcohol)
Quick Guide:
1. Swab the top of your sterile vial with alcohol prep pad (70% isopropyl alcohol)
2. Mix 7.5mL distilled water with 1.0mL vinegar
3. Add Whatman syringe filter
4. Add sterile ~20ga. needle to end of Whatman filter
5. Inject the 8.5mL of solution into the sterile vial
6. You now have sterile 0.6% acetic acid
Detailed Directions:
1. Wash you hands thoroughly
2. Optional: wear alcohol treated exam gloves (rub your gloved hands together with 70% isopropyl alcohol on them until dry)
3. Using a sterile alcohol prep pad, swab the top of your sterile glass vial (into which the acetic acid solution will be held in)
4. Using the 10mL syringe with a ~20ga. needle on the end, draw up 7.5mL distilled water
5. Using the same syringe, now draw up 1mL vinegar
6. Remove needle from the syringe and discard
7. Attach 0.2-0.22um Whatman sterile syringe filter (do not touch the free end that will have a needle on it)
8. Put a new, sterile needle (~20 gauge) onto the free end of the Whatman filter (do not touch needle)
a. Do not use the same needle on the Whatman that was used to originally draw up the unsterile vinegar and distilled water.
9. Put a ~20 gauge sterile needle into the top of your sterile glass vial to act as a vent
10. Inject the acetic acid solution into the vial
11. You are now done and should have sterile 0.6% acetic acid
Notes:
1. These items MUST be sterile: 20-22ga. Needles, whatman filter, glass vial
2. Whatman filter: These small, sterile filters are used to filter the acetic acid solution so it is sterile. It does not matter that the liquid in your syringe (distilled water & vinegar) is not sterile, nor does it matter that the syringe itself is not sterile. Once the liquid goes through the filter it is STERILE. Thus, everything after the filter must be sterile!
3. You will most likely use 1mL (milliliter) of 0.6% AA to reconstitute your IGF-1. Thus, you should make at least 1.5mL. In reality, it***8217;s just as easy to make 8.5mL as I have stated in the above directions. You will have plenty for use later then.
4. Do NOT reuse the Whatman filter nor any needles! Discard immediately.
Reconstituting IGF-1lr3
Reconstitution is simply the addition of the 0.6% AA to your lyophilized IGF-1.
Assumption: 1mg/mL IGF-1/AA (1mg IGF-1 will be combined with 1mL AA; 1mg IGF-1 is the same as 1,000mcg)
1. Swab the top of your IGF-1 vial with a sterile alcohol prep pad
2. Swab the top of your 0.6% AA vial with a sterile alcohol prep pad
3. Using either multiple insulin syringe volumes (example: 2 x 0.5cc) or a single large syringe, obtain 1.0mL of 0.6% AA.
4. In the IGF-1 vial, insert a sterile ~20 ga. needle to act as a vent
5. Inject the 1.0mL of AA very slowly and dribble it down the side of the vial.
a. Be very careful with this peptide as it is very delicate!
6. Remove the needle & syringe and discard
7. Gently swirl the vial or roll between your hands.
a. Again, be very gentle here
8. You now have 1mg/mL of IGF-1
a. This is the same as: 1,000mcg/mL
Notes:
1. If you added 2mL of AA, it would be a 0.5mg/mL
