The body's natural healing peptide that accelerates tissue repair and reduces inflammation
Written by Michael Carroll — Owner, Director of Research · Reviewed by the Peptide Initiative Research Team · Editorial standards
Subcutaneous
Route
4 recommended
Sites
Twice weekly
Frequency
Preparation
Bacteriostatic water or normal saline
Sterile syringe (3-5 mL recommended)
Sterile needle (25-27 gauge)
Alcohol swabs
Vial of Thymosin Beta-4 powder
Pro tip
Prepare all supplies on a clean surface before you begin. Having everything ready makes the process smoother and more sterile.
Mixing
Wash your hands thoroughly with soap and water. Gather all supplies on a clean, flat surface.
Remove the plastic cap from the peptide vial and wipe the rubber stopper with an alcohol swab. Let it air dry.
Draw the appropriate amount of bacteriostatic water into a sterile syringe.
Insert the needle into the vial at an angle, aiming at the inside wall of the vial. Slowly push the plunger to let the water trickle down the glass wall -- do NOT squirt directly onto the powder.
Once all water is added, gently swirl the vial in a slow circular motion. Never shake the vial, as this can damage the peptide bonds.
Continue swirling until the powder is completely dissolved and the solution is clear. If particles remain, let the vial sit for a few minutes and swirl again.
Label the vial with the date of reconstitution, the peptide name, and the concentration (e.g. 250mcg per 0.1mL).
Example calculation
For a 5 mg vial: Add 5 mL bacteriostatic water = 1 mg/mL solution. Each 0.1 mL = 0.1 mg (100 mcg)
Dose calculation
Desired dose (mg) × mL per mg concentration = volume to inject. Example: 2 mg dose from 1 mg/mL solution = 2 mL
Pro tip
Always add the bacteriostatic water slowly, letting it run down the side of the vial. Never shake the vial -- swirl gently to avoid damaging the peptide.
Location
Site 01
Abdomen
Pinch the skin 2 inches from navel. Avoid the area directly around the belly button. Rotate between left and right sides.
Site 02
Outer Thigh
Middle third of the outer thigh. Keep at least 4 inches above the knee and below the hip. Alternate legs each injection.
Site 03
Upper Arm
Back or outer area of the upper arm. This site may require assistance from another person for proper technique.
Site 04
Rotate sites with each injection
Follow your provider's instructions for this injection site. Rotate sites regularly to prevent tissue damage.
Rotate between 4 sites to prevent tissue buildup and ensure consistent absorption.
Pro tip
Rotate your injection sites with each dose to prevent lipohypertrophy (buildup of fatty tissue). Keep a simple log of where you last injected.
Step by step
Clean skin with alcohol swab and let dry
Pinch skin gently to create subcutaneous fold
Insert needle at 45-90 degree angle
Inject slowly over 2-3 seconds
Withdraw needle and apply light pressure
Rotate injection sites to prevent lipohypertrophy
Pro tip
This peptide uses subcutaneous. Inject at a 45-90 degree angle into pinched skin. Aspirate before injecting to ensure you haven't hit a blood vessel.
Timing
Optimal timing
Best time
Morning or evening (flexible)
With food?
Not affected by food intake
Stacking notes
Best administered on different days than other injectable peptides to minimize injection site stress
Sample daily schedule
Monday morning
2-5 mg injection
Site: Lower abdomen or outer thigh
First dose of the week (loading phase)
Thursday morning
2-5 mg injection
Site: Opposite thigh or upper arm
Second dose of the week (loading phase, then transition to maintenance)
Weekly (maintenance)
2 mg injection
Site: Rotate between abdomen, thighs, and arms
After 4-6 week loading phase, reduce to once weekly for maintenance
Dosing tiers
Dose
2-5 mg (community injectable practice for 'TB-500')
Frequency
Once or twice weekly
Duration
4-6 week loading phase, then weekly maintenance
Documented community/research-practice injectable dosing for tissue repair; thymosin beta-4 has ~100% bioavailability by the subcutaneous route [1]. Not an FDA-approved use.
Dose
0.1% (1 mg/mL) RGN-259 solution, 1 drop
Frequency
Several times daily (e.g. up to 5x/day)
Duration
4 weeks
0.1% RGN-259 ophthalmic solution used in phase III neurotrophic keratopathy and dry-eye trials, applied as eye drops several times daily for ~4 weeks [5].
Dose
450-1200 mg
Frequency
IV push daily for the first 3 days, then weekly
Duration
Total of 7 doses over ~5 weeks
Doses studied in the RGN-352 acute myocardial infarction clinical program, given IV after PCI [6].
Preservation
Before mixing
Keep in original vial at 2-8°C. Protect from light and freezing. Stable for 36+ months when properly stored
After mixing
Store at 2-8°C in refrigerator. Do not freeze reconstituted solution
Shelf life after mixing
14 days
Signs of degradation
Discard the vial immediately if you notice any of these:
Cloudiness or particles in solution
Discoloration (should be clear to slightly yellow)
Unusual odor
Solution has been at room temperature for >4 hours
Important
When to stop
Severe allergic reaction or anaphylaxis
Persistent injection site infection
Significant systemic adverse effects
Pregnancy is confirmed
No improvement in healing after 12 weeks (reassess approach)
Active systemic infection develops without antibiotic coverage
This is general educational information. Always consult with a healthcare provider before starting, stopping, or modifying any peptide therapy. Do not self-diagnose or treat serious medical conditions.
Clean technique checklist
Wash hands thoroughly with soap and water before handling supplies
Swab vial tops and injection site with alcohol and let dry
Never touch the needle tip or allow it to contact non-sterile surfaces
Use a new syringe and needle for each injection
Dispose of used sharps in a proper sharps container
Store reconstituted peptides according to the storage instructions above
Published research
Goldstein AL, Hannappel E, Sosne G, et al. · 2012
This study investigated the properties and effects of Thymosin Beta-4, contributing to our understanding of its mechanism of action and potential therapeutic applications.
Philp D, Kleinman HK · 2010
This study investigated the properties and effects of Thymosin Beta-4, contributing to our understanding of its mechanism of action and potential therapeutic applications.
Shrivastava S, Srivastava D, Olson EN, et al. · 2010
This study investigated the properties and effects of Thymosin Beta-4, contributing to our understanding of its mechanism of action and potential therapeutic applications.
Badamchian M, Fagarasan MO, Danner RL, et al. · 2003
This study investigated the properties and effects of Thymosin Beta-4, contributing to our understanding of its mechanism of action and potential therapeutic applications.
Sosne G, Ousler GW, et al. · 2022
RegeneRx Biopharmaceuticals (NCT01311518) · 2011
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