Cationic Ovalbumin (cOVA), Lyophilized

Cationic Ovalbumin (cOVA), Lyophilized - Cat Number: 9071 From Chondrex.

Research Field: Immunology, Allergy

Clonality: N/A

Cross-Reactivity:

Host Origin: N/A

Applications: N/A

Isotype: N/A

Detection Range: N/A

Sample Type: N/A

Concentration: N/A

Immunogen:

PRODUCT SPECIFICATIONS

DESCRIPTION: A cationized carrier protein for peptide conjugation
Peptides (less than 10 kDa) generally have very low antigenicity due to poor recognition by
antigen presentation cells (APCs). In order to increase antigenicity, peptides should be
conjugated to large molecule carrier proteins such as the widely used bovine serum albumin
(BSA) or keyhole limpet protein. For anti-peptide antibody production, cationic carrier proteins
are beneficial over typical carrier proteins for the following reasons: 1) cationic proteins can hold
more peptides because they contain more structural primary amines which generally work as
conjugation sites for peptides, and 2) positively charged cationic proteins can be selectively
processed as a presented antigen because of their high affinity for the negatively charged cell
surfaces of APCs. As carrier proteins with high antigenicity and peptide conjugation efficiency,
cationized BSA (cBSA) (Cat # 9070) and cationized ovalbumin (cOVA) (Cat # 9071) were
prepared by substituting negatively charged carboxyl groups with positively charged primary
amines.
APPLICATION: Use for peptide conjugation
QUANTITY: 10 mg
FORM: Lyophilized (no buffer components)
PURITY: >95%
ISOELECTRIC POINT: >10
CONJUGATION PROTOCOL: A. CARRIER PROTEIN ACTIVATION:
1. Dissolve 10 mg of cBSA or cOVA with 1 ml of PBS
2. Mix with 0.1 ml of sulfosuccinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate
(Sulfo-SMCC) (10 mg/ml in 0.05M phosphate buffered saline)
3. Incubate at room temperature for 1 hour
4. De-salt using gel filtration chromatography with PBS (e.g. Sephadex G-10)
5. Pool fractions collected from the first peak

B. PEPTIDE COUPLING:
1. Mix 1 ml of the pooled fraction with 1 ml of a sulfhydryl-containing peptide (10 mg/ml in
0.05M phosphate buffered saline).
NOTE: If the peptide is insoluble in 0.05M phosphate buffered saline, use a 6M
Guanidine/0.1M Phosphate buffer pH 7.4 or 10-20% DMSO in 0.05M phosphate buffered
saline.
2. Incubate at room temperature for 2 hours
3. Dialyze against 0.1M PBS pH 7.4 at 4°C
STORAGE: 4°C
STABILITY: 1 year
NOTES: Reagents for Immunizations:
1. Complete Freund’s Adjuvant, 1 mg/ml (Cat # 7008)
2. Complete Freund’s Adjuvant, 2 mg/ml (Cat # 7009)
3. Complete Freund’s Adjuvant, 4 mg/ml (Cat # 7001)
4. Incomplete Freund’s Adjuvant (Cat # 7002)
REFERENCES: N/A