antibody

Anti-Rainbow Trout Ig (H), Purified, (Clone IPA2C7) (mouse IgG1)

MBS520483

0.25 mg

375 USD

monoclonal

mouse

nonconjugated

IPA2C7

Antibody

Anti-Rainbow Trout Ig (H), Purified, (Clone IPA2C7) (mouse IgG1)

Rainbow Trout Ig (H)

Rainbow Trout Ig (H), Purified, (Clone IPA2C7) (mouse IgG1); Purified Anti-Rainbow Trout Ig Monoclonal Antibody

Monoclonal

Mouse IgG1

IPA2C7

Mouse

Rainbow Trout

Trout lg (H) (Immunoglobin heavy chain in Rainbow trout species)

Purified

Antibody Concentration: 1.0 mg/ml

Stable at 4 degree C. For long term storage, aliquot and freeze unused portions at -20 degree C in volumes appropriate for single usage. Avoid repeated freeze/thaw cycles.

Immunogen: Immunogen: Purified Rainbow trout Ig (heavy and light chains). Fusion Partner: X63Ag8.653 myloma cells

Presentation: Purified Ig buffered in PBS with the addition of 0.02% NaN3 (PEG Purified from Ascitic Fluid). Method: ELISA METHOD. I. Prepare a stock solution of 10-20 flg/ml of virus antigens in carbonate coating buffer, pH 9.6 and add 100fll/well (ie 1.0-2.0flg of antigen/well). Incubate at 4°C overnight. (For bacteria, dilute 1.0 00 660 nm of suspension 1/100 in water, add 100 fll per well and dry down overnight at 37°C. If your antigen is fish serum, you must dilute I/IO-I/SO in carbonate buffer and add 100 fll per well.) . 2. The next day, rinse plate 3 times quickly with PBS-0.05% tween 20 detergent, pH 7.4. 3. Block the plate (100 fll per well) with PBS, pH 7.4, containing 1-2% pre-immune goat serum. Incubate for I hour at room temperature. 4. Flick out the blocking buffer and rinse plate 3 times quickly with PBS-0.05% tween 20. 5. Add 100 fll per well of the salmon immune serum, diluted I/IO-I/SO in *PBS-0.05% tween 20. Incubate for 2 hours at room temperature or overnight at 4°C for best results. 6. Wash plate 3 times with PBS-0.05% tween 20 quickly. Then wash 3 x 10 minutes with PBS-0.05% tween 20 (ie. fill up the wells and let plate sit for 10 minutes, repeat 2 times) This longer washing step will reduce background. 7. Do not let the plate sit dry for long before adding III 00-1 /40M dilution ofCLFOOIAP in *PBS-0.05% tween 20 to all the wells. Incubate for 2 hours at room temperature. 8. Wash as in step 6. 9. Do not let the plate sit dry long before adding 100 fll per well of goat anti-mouse IgG I-Alk. Phos. secondary antibody:~, diluted 111 000-1/2000 in PBS-0.05% tween 20-0.5% pre-immune goat serum. Incubate for 2 hours at room temperature or I hour at 37°C. 10. Again, wash as in step 6. II. Add 100 ul per well of A Ik. Phos. substrate (one Sigma 104-105 Alk. Phos. substrate tablet in 5ml Diethanolamine buffer, pH 9.6). Incubate for 30-65 minutes in the dark. Read plate at 405 nm. *If background IS high, add 0.5% pre-Immune goat serum to the buffer solution. For optimal results in various applications, it is recommended that each investigator determine dilutions appropriate for individual use. In ELISA and Western blot assays, the recommended dilution for maximum activity is 1/100. It is recommended that the goat anti-mouse IgG 1 Alk. Phos. secondary antibody is used as this will increase the sensitivity 3-5 times above a normal multi-valent secondary antibody.

The anti-Rainbow trout Ig monoclonal antibody can be used to indirectly detect a generalized or pathogen-specific increase in immunoglobulin level. This monoclonal antibody only recognizes the immunoglobin heavy chain found in Oncorhynchus species (ie. Coho, Chum, Sockeye and Chinook) but not the Atlantic salmon heavy chain. This antibody can be used as a detection reagent for fish infection by either detecting a generalized increase in fish immunoglobulin level or by detecting specific antibody. For specific antibody detection, known infectious agents are used as the antigen in an ELISA assay, followed by samples of infected fish serum and then the anti-Rainbow Trout Ig monoclonal antibody. See ELISA protocol below. Specific uses for this monoclonal antibody include ELISA, Western blots and immunofluorescence (ie. FACS).

0.25 mg

375 USD