Antibody, rabbit

Kato, K., Hamaguchi, Y., Fukui, H., and Ishikawa, E. (1975b) Enzyme-linked immunoassay. II. A simple method for synthesis of the rabbit antibody-b-D-galactosidase complex and its general applicability. J. Biochem. (Tokyo) 78, 423. 

Mandy, W.J., Rivers, M.M., and Nisonoff, A. (1961) Recombination of univalent subunits derived from rabbit antibody./. Biol. Chem. 236, 3221. 

Palmer, J.L., and Nissonoff, A. (1963) Reduction and reoxidation of a critical disulfide bond in the rabbit antibody molecule./. Biol. Chem. 238, 2393-2398. 

Yoshitake, S., Yamada, Y., Ishikawa, E., and Masseyeff, R. (1979) Conjugation of glucose oxidase from Aspergillus niger and rabbit antibodies using N-hydroxysuccinimide ester of N-(4-carboxycyclohexyl methyljmaleimide. Eur.J. Biochem. 101, 395-399. 

An example of an evanescent wave fiber optic immunoassay and the associated optics has been described in detail for measurement of anti-rabbit IgG.(130) Rabbit antibody is immobilized on the distal face of an optical fiber. Unlabeled anti-rabbit IgG competes with fluorescein-labeled anti-rabbit IgG for rabbit antibody binding sites... 

Caspase-3 Activation Analysis fixed cells were washed and incubated overnight with rabbit anti-active caspase-3 monoclonal antibody followed by FITC goat anti-rabbit antibody. Then cells were washed and mixed with PI and analyzed by BD FACSCalibur flow cytometer. For WB, cells are lysed with SDS and proteins were analyzed with caspase 3 Asp-175, p53 and yH2AX Ser-139. 

Add the primary rabbit antibody directed against the antigen desired to the slide, making sure to cover the entire specimen. Work quickly to avoid any drying. Cover the chamber and incubate for 30 min (see Note 5). 

Apply enzyme-conjugated anti-species antibody onto the slide and incubate for approximately 15 min in a humidity chamber/box at room temperature. If the primary antibody is a rabbit antibody, an enzyme-conjugated anti-rabbit antibody is applied. 

Common haptens used for labeling DNA probes for BISH assays are biotin, DIG, DNP, FITC, and Texas Red. Based on the size of your DNA targets, you may choose from a direct detection or an indirect detection for BISH assays. In general, an indirect detection system can provide better sensitivity compared to a direct detection system. For an indirect detection, you need to select a combination of two antibodies raised with two different animal species, such as a mouse anti-DIG antibody and a rabbit anti-DNP antibody, so that enzyme-labeled anti-mouse antibody and anti-rabbit antibody can be applied for signal detection. If a direct BISH detection is going to be applied, anti-hapten antibodies raised in the same animal species that are labeled with either AP or HRP enzyme molecules... 

Cy3-conjugated goat-anti-mouse and Cy3-conjugated goat-anti-rabbit antibodies (Jackson Immunoresearch). 

Secondary antibody. For example, anti-rabbit antibody. 

Fig. 5. (a) Layout of a slide that was spotted and immobilized using the anti goat IgG rabbit antibody and the anti rabbit IgG goat-antibody, (b) Fluorescent image after incubation of a Cy5-labeled goat IgG. (c) Fluorescent image after incubation of a Cy5-labeled rabbit IgG. 

To each filter, add 5 mL of Tris/saline/BSA and the appropriate dilution of swine anti rabbit antibody (usually about L300) to each filter, and incubate for 30 min at room temperature on a rocking platform. 

AMOUNT OF PASSIVELY TRANSFERRED RABBIT ANTIBODIES ( ELISA end-point )... 

Eichmann, K. Kindt, T.J. (1971). The inheritance if individual antigenic specificities of rabbit antibodies to streptococcal carbohydrates. J. Exp. Med. 134,532-552. 

Sections (4 xm thick) of formalin-fixed and paraffin-embedded tissues are deposited on a coated slide, deparaffinized, rehydrated, and rinsed in PBS. They are placed in sodium citrate buffer-containing plastic jars and heated twice for 5 min each in a microwave oven. Following cooling at room temperature for 20 min, the sections are treated with 0.3% trypsin. They are washed in PBS, blocked with normal serum for 10 min, and then overlaid with fluorescein-conjugated rabbit antibodies to human IgG at 1 20 dilution in PBS for 45 min at room temperature. 

Fig. 18.3. Schematic representation of the photoaffinity immobilization of the antibody cholera toxin (anti-CT). After irradiation of the photobiotin layered electrode in presence of the antibody, a covalent bond has been formed between the electrode and the antibody keeping its accessibility for immunoreactions with HRP-IgG anti-rabbit antibody.

Cholera toxin B subunit-biotin labeled (lyophilized powder, biotin content 0.9mol/mol protein), peroxidase-labeled IgG anti-rabbit antibody (HRP-Ab, from goat, protein content 0.8mg/ml, affinity isolated antibody), anti-cholera toxin (from rabbit, protein content 48mg/ml, purified toxin from Vibrio cholerae), biotin monoclonal anti-rabbit IgG -y-chain specific (from mouse, protein content 4.2mg/ml), glucose oxidase-biotinamidocaproyl labeled (GOX-B, from Aspergillus niger, lyophilized powder containing 40-70% protein, 137 U/mg), polyoxyeth-ylene-sorbitan monolaurate (Tween 20), bovine serum albumin (fraction... 

Fig. 26.2. Amperometric immunosensors set-up using a biotinylated copolymer poly(pyrrole-biotin, pyrrole-lactitob-ionamide) coated platinum or glassy carbon electrodes and three enzymatic markers (GOX-B, PPO-B, HRP-Ab) for the detection of cholera antitoxin. (A) HRP-immunosensor, (B) GOX-B-immunosensor, (C) PPO-B-immunosensor. Mred/Mox = hydroquinone/quinone Gox = biotinylated glucose oxidase PPO — biotinylated polyphenol oxidase HRP-Ab = peroxidase-labeled IgG anti-rabbit antibody.

The rabbit antibodies used for capturing MAbs may also interact with some phage from the library. This might result in selection of many non-MAb specific clones. Thus, the library should be pre-incubated... 

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