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AbFlex AM-Tag antibody (rAb)

Host / Isotype

Mouse / IgG2a

Reactivity

Human, Not Species Specific

Applications

ChIP, ELISA, WB

Activ Motif logo

Cat No : 91111,91112 91111

Synonyms

H2av, histone, histone variant, h2az, h2a.z, drosophila, fruitfly, chromatin, H2AFV, drosophila h2az, antibody, antibodies, chromatin immunoprecipitation, chip, western blotting, wb, recombinant, rAb, rec Ab, AbFlex

Validation Data Gallery

  • AbFlex AM-Tag antibody (rAb) tested by ChIP. Active Motif’s pAM_1C_JunD Vector (Catalog No. 53044) was transiently transfected or mock transfected into HCT116 cells. Chromatin was harvested according to the instructions in the Tag-ChIP-IT Kit (Cat. No. 53022). 10 ug of the AM-Tag antibody was used to immunoprecipitate the cross-linked AM-Tag-JunD fusion protein. qPCR data shows enrichment of AM-Tag-JunD with the FOXRED qPCR primer set and little to no enrichment in the mock transfections or when using the negative control Unt12 qPCR primer set.

    AbFlex AM-Tag antibody (rAb) tested by ChIP. Active Motif’s pAM_1C_JunD Vector (Catalog No. 53044) was transiently transfected or mock transfected into HCT116 cells. Chromatin was harvested according to the instructions in the Tag-ChIP-IT Kit (Cat. No. 53022). 10 ug of the AM-Tag antibody was used to immunoprecipitate the cross-linked AM-Tag-JunD fusion protein. qPCR data shows enrichment of AM-Tag-JunD with the FOXRED qPCR primer set and little to no enrichment in the mock transfections or when using the negative control Unt12 qPCR primer set.

  • AbFlex AM-Tag antibody (rAb) tested by Western Blot. Active Motif’s pAM_1C_JunD Vector (Catalog No. 53044) was transfected or mock transfected into HCT116 cells. 48 hours post-transfection nuclear lysates were prepared. 15 ug lysate was loaded per well. Western blot was performed using 1 ug/mL of AM-Tag antibody.

    AbFlex AM-Tag antibody (rAb) tested by Western Blot. Active Motif’s pAM_1C_JunD Vector (Catalog No. 53044) was transfected or mock transfected into HCT116 cells. 48 hours post-transfection nuclear lysates were prepared. 15 ug lysate was loaded per well. Western blot was performed using 1 ug/mL of AM-Tag antibody.

Product Information

Tested Applications ChIP, ELISA, WB

Applications Validated by Active Motif: ChIP: 10 ug per ChIP WB: 0.5 - 2 ug/ml dilution Bead-based ELISA: 7 - 560 ng/ml
Tested Reactivity Human, Not Species Specific
Host / Isotype Mouse / IgG2a
Class Recombinant
Type Antibody
ModificationAM-Tag
Immunogen This antibody was raised against Active Motif's unique AM-Tag sequence for use with Tag-ChIP-IT (Cat No. 53022) analysis.
Full Name AbFlex AM-Tag antibody (rAb)
Synonyms H2av, histone, histone variant, h2az, h2a.z, drosophila, fruitfly, chromatin, H2AFV, drosophila h2az, antibody, antibodies, chromatin immunoprecipitation, chip, western blotting, wb, recombinant, rAb, rec Ab, AbFlex
Molecular weight 15 kDa
GenBank accession numberNP_524519
RRIDAB_2793779
Purification Method Protein A Chromatography
Buffer Purified IgG in 200 mM Hepes pH 7.5, 100 mm NaCl, 50 mM NaOAc, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.
Storage Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Background Information

AbFlex antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis Tag, a Biotinylation Tag for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports. AbFlex AM-Tag antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells. The AM-Tag antibody is specifically designed for use with Active Motif’s unique AM-Tag sequence in combination with the Tag-ChIP-IT Kit (Catalog No. 53022). Simply clone your protein of interest into Active Motif’s pAM_1C Empty Vector (Catalog No. 53023), or add the AM-Tag sequence to the C-terminus of your protein expression vector. Following transfection, cell lysates are prepared and screened by Western blot for expression of the AM-Tag fusion protein using the AM-Tag antibody. Optimization may be required to identify the optimal conditions for transfection and protein expression. Once transfection conditions have been optimized, the Tag-ChIP-IT Kit can be used to isolate chromatin and perform ChIP. The AM-Tag has minimal cross reactivity with mammalian samples to ensure low background signal. The tag is also unstructured, which allows the tag to protrude from the protein of interest for maximum exposure during immunoprecipitation. This increases the enrichment efficiency of low abundance transcription factors for more reliable and consistent ChIP results. Use of the AM-Tag for chromatin immunoprecipitation avoids the need for protein-specific ChIP-validated antibodies. It also enables the analysis of sequence variants, mutations and truncations on gene regulation.