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Antibodies

AbFlex® AM-Tag antibody (rAb) (biotin)

Catalog No: 91109 Format: 100 µg $405 Buy Now
Catalog No: 91110 Format: 10 µg $85 Buy Now

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Antibody Type:Recombinant
Isotype:IgG2a
Purification:Ni-NTA
Host:Mouse
Molecular Weight:15 kDa
Reactivity:Other (Wide Range)

Applications

ChIP Validated Western Blot Validated Enzyme-linked Immunosorbent Assay Validated

for AbFlex AM-Tag antibody (rAb) (biotin)Application Notes


Validated Applications:
ChIP: 10 µg per ChIP
WB: 0.5 - 2 µg/ml dilution
Bead-based ELISA: 7 - 560 ng/ml

for AbFlex AM-Tag antibody (rAb) (biotin)Immunogen

This antibody was raised against Active Motif's unique AM-Tag sequence for use with Tag-ChIP-IT® (Cat No. 53022) analysis.

for AbFlex AM-Tag antibody (rAb) (biotin)Buffer

Purified IgG in 50mM sodium phosphate pH 8.0, 150mM NaCl, and 100mM imidazole with 30% glycerol and 0.035% sodium azide.

 
AbFlex<sup>®</sup> AM-Tag antibody (rAb) (biotin) tested by ChIP.

AbFlex® AM-Tag antibody (rAb) tested by ChIP.
Active Motif’s pAM_1C_JunD Vector (Catalog No. 53044) was transiently transfected, mock transfected or transfected with pAM_1C Empty Vector (#53023) into HCT116 cells. Chromatin was harvested according to the instructions in the Tag-ChIP-IT® Kit (#53022). 10 µg 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, empty vector transfections, or when using the negative control Unt12 qPCR primer set.

AbFlex<sup>®</sup> AM-Tag antibody (rAb) (biotin) tested by Western blot.

AbFlex® AM-Tag antibody (rAb) tested by Western Blot.
Active Motif’s pAM_1C_JunD Vector (Catalog No. 53044) was transfected, mock transfected or transfected with pAM_1C Empty Vector (Catalog No. 53023) into HeLa or HCT116 cells using 10 µg DNA and 30 µl FuGENE transfection reagent, 48 hours post-transfection nuclear lysates were prepared. 20 µg lysate was loaded per well. Western blot was performed using a 1:500 dilution of AM-Tag antibody.

for AbFlex AM-Tag antibody (rAb) (biotin)Background

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, an avidin tag sequence 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® Histone H2Av antibody (biotin) was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells. The antibody was directly labeled with biotin using the biotin ligase, BirA.

The nucleosome is the smallest subunit of chromatin and consists of 147 base pairs of DNA wrapped around an octamer of core histone proteins (two each of Histone H2A, Histone H2B, Histone H3 and Histone H4). Histone H2A.Z (H2AZ, H2AFZ) is a histone H2A variant, a protein similar to canonical H2A but with different molecular identity and unique functions. H2A.Z is highly conserved during evolution. It plays an important role in basic cellular mechanisms such as gene activation, chromosome segregation, heterochromatic silencing and progression through the cell cycle.In Drosophila, the H2A variant corresponding to H2AZ is H2Av. H2Av is an essential protein in Drosophila and has been implicated in both activation and repression of transcription. H2Av is localized to centromeric heterochromatin in Drosophila and flies lacking H2Av have reduced levels of heterochromatin components at the centromeres. However, H2Av nucleosome distribution throughout the rest of the Drosophila genome correlates with genes that have an open and uniform chromatin architecture at promoter regions.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with AbFlex® AM-Tag antibody (rAb) (biotin).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use AbFlex® AM-Tag antibody (rAb) (biotin).

for AbFlex AM-Tag antibody (rAb) (biotin)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for AbFlex AM-Tag antibody (rAb) (biotin)Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting

Technical Data Sheet

AbFlex® AM-Tag antibody (rAb) (biotin)
 

Data Thumbnails

AbFlex<sup>®</sup> AM-Tag antibody (rAb) (biotin) tested by ChIP.

ChIP of AbFlex® AM-Tag antibody (rAb) (biotin).
(Click image to enlarge and see details.)

AbFlex<sup>®</sup> AM-Tag antibody (rAb) (biotin) tested by Western blot.

Western blot of AbFlex® AM-Tag antibody (rAb) (biotin).
(Click image to enlarge and see details.)