Mouse monoclonal antibody

Anti-Complement component C3a/C3a/(desArg)/C3 (human

CAT NO: GAU 013-16 | Application: ELISA, WB, IHC, IF

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Product Summary

Cat. No.

GAU 013-16

Specificity

GAU 013-16 is specific for human C3, C3a and C3a (desArg) (1) and does not cross-react with C4a or C5a. (2, 3)

Immunogen

Human C3a

Host

Mouse

Positive Species Reactivity

Human

Negative Species Reactivity

Not determined

Isotype

IgG1/k

Gene ID

718

Clone Number

K13/16-5.7

Epitope Specificity

GAU 013-16 recognizes a different epitope on the 9 kDa C3a than GAU 017-01 (2, 3). No reaction is seen with a synthetic octapeptide representing the C3a C-terminal. (2)

Unit Size

400 µL: Cat. No. GAU 013-16-04 1 mL: Cat. No. GAU 013-16-1 1 mg/mL +/- 15%. See Certificate of Analysis for details.

Purification

Protein A/G purified (from culture supernatant)

Form

Liquid

Solvent

0.01 M phosphate buffer, pH 7.4, containing 0.5 M NaCl and 15 mM sodium azide

Conjugation

Unconjugated BSA free

Storage

4-8ºC without exposure to light. No precautions necessary during handling.

Target

Complement C3a is an anaphylatoxin of 77 amino acid residues released by the action of the C3 convertases on the N-terminal of the alpha chain of C3. It is rapidly inactivated by serum carboxypeptidase N which removes the C-terminal arginine residue generating C3a (desArg).

Application

ELISA: GAU 013-16 can be used as a capture antibody in sandwich ELISA with GAU 017-01 as a biotinylated detection antibody. (4, 5) GAU 013-16 can also be used to immunopurify C3a (3). GAU 013-16 effectively inhibits the biological activity of C3a in a guinea-pig platelet activation assay. (2)

WB: GAU 013-16 was used in Western blotting. (2, 3, 5)

Immunoblot with GAU 013-16: Lane 1: Purified C3 Lane 2: Purifed C3 reduced/DTT Lane 3: Purified C3a Lane 4: EDTA-plasma Lane 5: Zymosan-activated serum

IHC: GAU 013-16 was used in IHC in formaline fixed paraffin embedded tissue with a dilution of 1:150.

IHC staining of human kidney with GAU 013-16. Strong staining of veins of nephritis kidney was seen. Dilution: 1:150

IHC staining of human liver with GAU 013-16. Strong staining of plasma of veins in liver tissue. Dilution: 1:150

IHC staining of human tonsil with GAU 013-16. Plasma in lumen of veins is stained strongly in tonsil. Dilution: 1:150

IF: GAU 103-16 was used in IF with a dilution of 1:75.

IF staining in human kidney with GAU 013-16. Strong staining of plasma of nephritis kidney was seen. Dilution: 1:75

Scientific References

1. Oppermann M, Haubitz M, Quintin E, Götze O (1988) Complement activation in patients with renal failure as detected through the quantitation of fragments of the complement proteins C3, C5, and Factor B. Klin Wochenschr 66:857-864.

2. Nezlin R, Freywald A, Oppermann M (1993) Proteins separeted from human IgG molecules. Mol. Immunol. 30:935-940.

3. Puschel GP, Oppermann M, Muschol W, Gotze O, Jungermann K. (1989) Increase of glucose and lactate output and decrease of flow by human anaphylatoxin C3a but not C5a in perfused rat liver. FEBS Lett. 16;243(1):83-7.

4. Khodoun M, Strait R, Orekov T, Hogan S, Karasuyama H, Herbert DBR, Köhl J, Finkelman FD (2009) Peanuts can contribute to anaphylactic shock by activating complement. J Allergy Clin Immunol 123:342-351.

5. Thomas SN, van der Vlies AJ, O’Neil CP, Reddy ST, Yu SS, Giorgio TD, Swartz MA, Hubbell JA (2011) Engineering complement activation on polypropylene sulfide vaccine nanoparticles. Biomaterials 32: 2194-2203.