1Mouse CIA – A Visual Guide
For Research Use Only - Not Human or Therapeutic Use

MICE: CHOOSE 1 STRAIN
DBA/1 (DBA) mice OR C57BL/6 (B6) mice•••

7-8 weeks old
SPF housing conditions
High fat diet

RECOMMENDED ADJUVANTS & COLLAGENS: CHOOSE 1 ADJUVANT + 1 COLLAGEN
Catalog #Adjuvants

Catalog #Collagens

7002

Incomplete Freund’s Adjuvant (IFA) (DBA - Booster only)

20011

Chick Type II Collagen, Lyophilized (DBA or B6)

7008

Complete Freund’s Adjuvant (CFA), 1 mg/ml (DBA)

20012

Chick Type II Collagen, 2 mg/ml Solution (DBA or B6)

7001

Complete Freund’s Adjuvant (CFA), 4 mg/ml (DBA)

20021

Bovine Type II Collagen, Lyophilized (DBA)

7023

Complete Freund’s Adjuvant (CFA), 5 mg/ml (B6)20022

Bovine Type II Collagen, 2 mg/ml Solution (DBA)

IMMUNIZATION SCHEDULE: CHOOSE 1 OPTIONOPTION 1: Single Immunization Protocol (DBA mice)
Day 0: Subcutaneous injection: Collagen emulsion with CFA: 4 mg/ml M. tuberculosis
Day 0 - 28: Score arthritis every 7 days
Day 28 - 56: Score arthritis every 2-3 days: Arthritis incidence 90 - 100%Day 56: End of experiment.
OROPTION 2: Booster Immunization Protocol
Day 0: Subcutaneous injection: Collagen emulsion with CFA: 1 mg/ml (DBA) or 5 mg/ml M. tuberculosis (B6)
Day 0 - 28: Score arthritis every 7 days
Day 21: Subcutaneous injection (booster): Collagen emulsion with IFA (DBA) or CFA: 5 mg/ml M. tuberculosis (B6)
Day 28 - 56: Score arthritis every 2-3 days: Arthritis incidence 90 - 100% (DBA), 50-70% (B6)
Day 56: End of experiment© 2020 Chondrex, Inc. All Rights Reserved, MCIA VG 2.0
16928 Woodinville-Redmond Rd NE Suite B-101
Woodinville, WA 98072

Phone: 425.702.6365 or 888.246.6373
Fax: 425.882.3094

www.chondrex.com
support@chondrex.com

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PREPARE EMULSION WITH A HOMOGENIZER: DO NOT USE THE DOUBLE-SYRINGE METHOD!
Figure 1

Figure 2

Figure 3

Collagen

CFA
Procedure*:
1. Gather materials (Figure 1. Homogenizer blade (a), 10 ml syringe with a 3-way stopcock (b), and 1 ml Hamilton syringe (c)).
2. Lock stopcock.
3. Keep setup on ice (Figure 2).
4. Add CFA to syringe.
5. Add an equal volume of collagen solution, drop-wise (Figure 3).
6. Mix at 10,000 - 30,000 rpm for 2 minutes. Move the syringe up and down while mixing.
7. Cool syringe on ice for 5 minutes.
8. Mix at 10,000 - 30,000 rpm for 2 minutes. Move the syringe up and down while mixing.
9. Evaluate emulsion with the drop test (next page).
10. Transfer the emulsion to a Hamilton syringe using the 3-way stopcock.

*For more information, please see Chondrex, Inc’s Mouse CIA Protocol© 2020 Chondrex, Inc. All Rights Reserved, MCIA VG 2.0
16928 Woodinville-Redmond Rd NE Suite B-101
Woodinville, WA 98072

Phone: 425.702.6365 or 888.246.6373
Fax: 425.882.3094

www.chondrex.com
support@chondrex.com

3EMULSION DROP TESTOREmulsion holds shape: Ready to use!
(Use immediately or store at 4⁰C up to 3 hours)

Emulsion disperses on water surface(Add one drop of adjuvant to the emulsion andhomogenize again)

IMMUNIZATION(a) Emulsion in Hamilton Syringe(no air bubbles!)(b) Needle (25 or 27-gauge x 5/8”)(c) Restraining Device© 2020 Chondrex, Inc. All Rights Reserved, MCIA VG 2.0
16928 Woodinville-Redmond Rd NE Suite B-101
Woodinville, WA 98072

Phone: 425.702.6365 or 888.246.6373
Fax: 425.882.3094

www.chondrex.com
support@chondrex.com

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SUBCUTANEOUS INJECTION: 0.1 ml per injection

AFTER INJECTION:(a)(b)

DO NOT USE
IP (a) OR IV (b)INJECTIONS TOIMMUNIZE MICE!

© 2020 Chondrex, Inc. All Rights Reserved, MCIA VG 2.0
16928 Woodinville-Redmond Rd NE Suite B-101
Woodinville, WA 98072

Phone: 425.702.6365 or 888.246.6373
Fax: 425.882.3094

www.chondrex.com
support@chondrex.com