DM4 Recombinant Rabbit Monoclonal Antibody [3H3]
Catalog# HA721153
DM4 Recombinant Rabbit Monoclonal Antibody [3H3]
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ELISA
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Species independent
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unconjugated
Overview
Product Name
DM4 Recombinant Rabbit Monoclonal Antibody [3H3]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
DM4 coupled with OVA.
Species Reactivity
Species independent
Validated Applications
ELISA
Conjugation
unconjugated
Clone Number
3H3
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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ELISA
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1:10,000
Target
Function
Ravtansine (DM4) is a maytansinoid, a chemical derivative of maytansine being investigated as the cytotoxic payload of a number of antibody-drug conjugates (ADCs). Microtubules are dynamic cytoskeletal polymers that switch stochastically between states of growing and shortening, called “dynamic instability”. They function in the precise segregation of chromosomes during cell division, transport of cellular cargos, and positioning and movement of intracellular organelles. Inhibition of microtubule function leads to cell cycle arrest and cell death. Microtubule-targeted drugs including the Vinca alkaloids, taxanes, and epothilones suppress the dynamic instability of microtubules, induce mitotic arrest, inhibit cell proliferation and induce apoptosis. The anticancer properties of maytansinoids have been attributed to their ability to disrupt microtubule function. The maytansinoid emtansine (DM1), for example, binds at the ends of microtubules and thereby suppress their dynamic instability. It is synthesized in order to link maytansinoids to antibodies via disulfide bonds. Maytansinoids inhibit tubulin polymerization and microtubule assembly and enhance microtubule destabilization, so there is potent suppression of microtubule dynamics resulting in a mitotic block and subsequent apoptotic cell death. DM4 can be used in the preparation of antibody drug conjugate. Although S-methyl DM1 and S-methyl DM4 inhibited microtubule assembly more weakly than maytansine, they suppressed dynamic instability more strongly than maytansine. Like vinblastine, the maytansinoids potently suppress microtubule dynamic instability by binding to a small number of high affinity sites, most likely at microtubule ends. Thus, the maytansine derivatives that result from cellular metabolism of the antibody conjugates are themselves potent microtubule poisons, interacting with microtubules as effectively as or more effectively than the parent molecule.
Background References
1. Smith SV. Technology evaluation: cantuzumab mertansine, ImmunoGen. Curr Opin Mol Ther. 2004;6:666–74.
2. Erickson HK, Widdison WC, Mayo MF, et al. Tumor delivery and in vivo processing of disulfide-linked and thioether-linked antibody-maytansinoid conjugates. Bioconjugate Chem. 2010;21:84–92.
Images
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Anti-DM4 Antibody (HA721153) in indirect ELISA.
Indirect ELISA analysis of DM4 was performed by coating wells of a 96-well plate with 50 µl per well of DM4-BSA diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 50 µl per well of a DM4 recombinant rabbit monoclonal antibody starting at a concentration of 10 µg/mL and serially diluting it to a concentration of 0.169 ng/mL for 2 hours at room temperature. The plate was washed and incubated with 50 µl per well of an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:15,000 for one hour at room temperature. Detection was performed using an Ultra TMB Substrate for 30 minutes at 37℃. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm. -
Competitive ELISA analysis of DM4 was performed by coating wells of a 96-well plate with 50 µl per well of DM4-OVA diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 1%BSA blocking buffer, Serially diluted DM4 monoclonal antibody (HA721153), ranging in concentration from 4,000 ng/mL to 3.9 ng/mL, was added to the wells along with DM4 or DM4-OVA at a final concentration of 10 µg/mL each. The plate was incubated at room temperature for 1 hour. The plate was washed and incubated with 50 µl per well of an HRP-conjugated goat anti-Rabbit IgG secondary antibody at a dilution of 1/15,000 for one hour at room temperature. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"