Mab-ZAP [IT-04, KIT-04]

a tool to “piggyback” onto YOUR antibody via affinity-purified goat anti-mouse IgG; targeting cells that recognize YOUR primary mouse monoclonal IgG antibody, eliminated via saporin

SKU: IT-04 Category: Quantity: Individual 25 ug, Individual 100 ug, Individual 250 ug, Individual 1 mg, Kit w/controls 25 ug, Kit w/controls 100 ug, Kit w/controls 250 ug, Kit (1 ab) w/controls & developing reagents, tests 1 ab, Kit (4 ab) w/controls & developing reagents, tests 4 abs, Kit (10 ab) w/controls & developing reagents, tests 10 abs | Antibody Type: affinity-purified | Host: goat | Reactivity: mouse | Conjugate: saporin | Usage: eliminates cells, screen antibodies |

Mab-ZAP uses your primary mouse monoclonal IgG antibody to target and eliminate cells that recognize your primary antibody. This secondary conjugate uses the secondary antibody (affinity-purified goat anti-mouse IgG) to “piggyback” onto YOUR mouse primary antibody. Mab-ZAP can be utilized for screening mouse IgG antibodies for internalization and/or their suitability to make potent immunotoxins.

Mab-ZAP is a chemical conjugate of affinity-purified goat anti-mouse IgG and the ribosome-inactivating protein, saporin. It uses your mouse primary antibody to target and eliminate cells. This secondary conjugate is used to evaluate the potential of a primary antibody to internalize.

Mab-ZAP is available individually (Cat. #IT-04) or as a kit (Cat. #KIT-04) which includes Mab-ZAPSaporin (Cat. #PR-01)Goat IgG-SAP (Cat. #IT-19) and reagents for developing a cytotoxicity assay.

Other ZAP Conjugates:

Need another ZAP Conjugate for your target? Check here for other species & targets including in vivo and in vitro options.

keywords: Mouse Antibody, Mouse Primary, Mouse Monoclonal, Anti-Mouse IgG, Screening, ADC, Antibody Drug Conjugate, in vitro, Secondary


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Saporin as a commercial reagent: its uses and unexpected impacts in the biological sciences-tools from the plant kingdom

Ancheta LR, Shramm PA, Bouajram R, Higgins D, Lappi DA (2022) Saporin as a commercial reagent: its uses and unexpected impacts in the biological sciences-tools from the plant kingdom. Toxins (Basel) 14(3):184. doi: 10.3390/toxins14030184

Read complete article.

Cellular uptake of vitamin B12: Role and fate of TCblR/CD320, the transcobalamin receptor.

Gick GG, Arora K, Sequeira JM, Nakayama Y, Lai SC, Quadros EV (2020) Cellular uptake of vitamin B12: Role and fate of TCblR/CD320, the transcobalamin receptor. Exp Cell Res 396(1):112256. doi: 10.1016/j.yexcr.2020.112256

Summary: The increased and sustained expression of TCblR in proliferating cells has been used to target toxins preferentially to cancer cells and can be potentially used for targeted delivery of other anti-cancer drugs. In 2010 the authors published a paper which evaluated the potential of using immunotoxins to eliminate cancer cells expressing TCblR the authors performed a series of in vitro experiments using their monoclonal antibody plus Mab-ZAP in varying concentrations. The results indicated that this is a viable therapeutic model that causes minimal peripheral damage.

Related Products: Mab-ZAP (Cat. #IT-04)

The EphA2 and cancer connection: potential for immune-based interventions

London M, Gallo E (2020) The EphA2 and cancer connection: potential for immune-based interventions. Mol Biol Rep 47(10):8037-8048. doi: 10.1007/s11033-020-05767-y

Summary: The authors review the most current mAb-based therapies against EphA2-expressing cancers currently in pre-clinical and/or clinical stages. They reference Sakamoto et al. who performed in vitro testing of two different EphA2 mAbs mixed with Mab-ZAP to discover their therapeutic potential against melanoma.

See: Sakamoto A et al. An Agonistic Antibody to EPHA2 Exhibits Antitumor Effects on Human Melanoma Cells. Anticancer Res 38:3273-3282, 2018.

Related Products: Mab-ZAP (Cat. #IT-04)

Targeting of embryonic annexin A2 expressed on ovarian and breast cancer by the novel monoclonal antibody 2448

Cua S, Tan HL, Fong WJ, Chin A, Lau A, Ding V, Song Z, Yang Y, Choo A (2018) Targeting of embryonic annexin A2 expressed on ovarian and breast cancer by the novel monoclonal antibody 2448. Oncotarget 9:13206-13221. doi: 10.18632/oncotarget.24152

Objective: To develop mAbs to potentially target oncofetal antigens and be repurposed for antibody or antibody drug conjugate (ADC) therapy.

Summary: The novel IgG1, 2448, was shown to target a unique glycosylated surface epitope on ANXA2. As a possible therapeutic candidate for ovarian and breast cancer, 2448 demonstrated anti-tumor activity via two independent mechanisms of action.

Usage: Cells were seeded in 96-well plates at 1000 or 2000 cells/well. Primary antibody, 2448 or ch2448 (10 μg/mL) was pre-mixed with appropriate secondary saporin conjugate, Mab-ZAP or Hum-ZAP.  The most significant decreases in cell viability (20% to 60%) were observed against the epithelial IGROV1 and MCF7 cell lines.  ATS created a Custom ADC by direct conjugation of saporin to ch2448 (ch2448-SAP).  As a control, an isotype chimeric IgG was also conjugated to saporin (IgG-SAP). Compared to using secondary saporin conjugates, ch2448-SAP induced and increase of  20–30% cytotoxicity.)

Related Products: Mab-ZAP (Cat. #IT-04), Hum-ZAP (Cat. #IT-22), Custom Conjugates

Evaluate Potential Targeting Molecules.

Kohls M (2006) Evaluate Potential Targeting Molecules. Nature Methods

Summary: Targeted toxins -- targeting agents conjugated to saporin -- are widely used to eliminate specific cell populations both in vitro and in vivo. For these molecules to be effective, it is vital that the targeting component of the conjugate specifically binds the cells of interest. A secondary conjugate, Streptavidin-ZAP, has been created by attaching the toxin saporin to streptavidin. The user can combine primary biotinylated material with Streptavidin-ZAP to quickly and economically screen potential targeting molecules for internalization and specificity. Once the appropriate targeting molecule is identified, a direct conjugation with saporin can be performed.

Related Products: Streptavidin-ZAP (Cat. #IT-27)

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Mab-ZAP: A tool for evaluating antibody efficacy for use in an immunotoxin.

Kohls MD, Lappi DA (2000) Mab-ZAP: A tool for evaluating antibody efficacy for use in an immunotoxin. BioTechniques 28(1):162-165. doi: 10.2144/00281pf01

Summary: Immunotoxins are useful tools for elimination of specific cell populations in vitro and in vivo for research and therapeutic applications. One of the factors limiting the use of immunotoxins is the selection of an appropriate antibody. Advanced Targeting Systems has created a reagent that allows researchers to select antibodies with the desired characteristics before an immuntoxin is made, purified, and assayed. Using a goat anti-murine IgG coupled to the ribosome-inactivating protein saporin, researchers can screen hundreds of antibodies in a time and cost-effective manner.

Related Products: Mab-ZAP (Cat. #IT-04), Rab-ZAP (Cat. #IT-05), Hum-ZAP (Cat. #IT-22), Rat-ZAP (Cat. #IT-26), Anti-M-ZAP (Cat. #IT-30), Goat-ZAP (Cat. #IT-36)

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Saporin (Cat. #PR-01)

Goat IgG-SAP (Cat. #IT-19)

Fab-ZAP mouse (Cat. #IT-48)

FabFc-ZAP mouse (Cat. #IT-89)

Anti-M-ZAP (Cat. #IT-30)

Choosing the Correct Secondary Conjugate

Higher concentration and less cytotoxicity

Assay Parameters for Mab-ZAP

Does Mab-ZAP bind heavy and light chain?

IgM Primary Antibody Assay

Mab-ZAP binds to Fc portion of mouse IgG

Secondary Conjugate Flow Cytometry

Sterilizing Primary Antibody

Cytotoxicity of Unbound Saporin

ZAP Antibody Internalization Kit Introduction

ZAP Antibody Internalization Kit Tutorial

ZAP Antibody Internalization Kit literature

Cytotoxicity Assay Protocol for ZAP Antibody Internalization Kits

Online Calculator: Preparing ZAP Antibody Kit Samples

Concentration Calculations: Convert molarity to mg/ml and mg/ml to molarity (PDF worksheet)

Preparing and Interpreting Cytotoxicity Data in vitro

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