Tested applications:WBIHC-PIF/ICCIPChIPChIP-seqRIPFCFC(Intra)ELISAMeDIPNucleotide ArrayDBFACSCoIPCUT&TagmeRIPInhibitionReactivity:Species independent
Western blot analysis of lysates from wild type (WT) and 293T cells transfected with GSK3B Protein, BCAT2-N Protein, BCAT2-C Protein, using DDDDK-Tag antibody (AE092) at 1:10000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.
Western blot analysis of lysates from wild type (WT), 293F transfected with COPB2-N Protein, COPB2-C Protein, using DDDDK-Tag antibody (AE092) at 1:10000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.
Chromatin immunoprecipitation was performed with 21.8 μg of cross-linked chromatin from 293T cells transfected with a GATA3 expression vector containing a single C-terminal DDDDK-Tag using 5 μg of Rabbit anti DDDDK-Tag (AE092). DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2 (RK20228). The ChIP sequencing results indicate the enrichment pattern of DDDDK-Tag across chromosome 2 (upper panel) and the genomic region encompassing SH3RF3, a representative gene enriched in DDDDK-Tag (lower panel).
Chromatin immunoprecipitation was performed with 21.8 μg of cross-linked chromatin from 293T cells transfected with a GATA3 expression vector containing a single C-terminal DDDDK-Tag using 5 μg of Rabbit anti DDDDK-Tag (AE092). DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2 (RK20228). The ChIP sequencing results indicate the enrichment pattern of DDDDK-Tag in the representative genomic region surrounding SH3RF3 gene.
Confocal imaging of 293F cells transfected with GFP-DDDDK-Tag (N) and 293F cells transfected with GFP-DDDDK-Tag (C) cells using DDDDK-Tag Rabbit mAb (AE092, dilution 1:1600) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.
Immunoprecipitation of DDDDK-Tag from 300 ug extracts of 293T cells transfected with a SERPINB1 expression vector containing a single N-terminal DDDDK-Tag was performed using 3 μg of DDDDK-Tag Rabbit mAb(AE092). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. The IP sample was eluted with 1X reducing Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using DDDDK-Tag Rabbit mAb (AE092) at a dilution of 1:2000.
Immunoprecipitation of DDDDK-Tag from 300 ug extracts of 293T cells transfected with a GSK3B expression vector containing a single C-terminal DDDDK-Tag was performed using 3 μg of DDDDK-Tag Rabbit mAb(AE092). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. The IP sample was eluted with 1X reducing Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using DDDDK-Tag Rabbit mAb (AE092) at a dilution of 1:2000.
Chromatin immunoprecipitation was performed with 20 μg of cross-linked chromatin from 293F cells (left) and 293F cells transfected with BATF3 (right), using 2 μg of DDDDK-Tag Rabbit mAb (AE092) and Rabbit IgG isotype control (AC042). The enrichment of immunoprecipitated DNA at different genomic loci was examined by quantitative PCR. The histogram compares the ratio of the immunoprecipitated DNA to the input at given loci.
Flow cytometry: 1X10^6 293T cells (negative control, left) and 293T (Transfection, right) cells were surface-stained with DDDDK-Tag Rabbit mAb (AE092, 2.5 μg/mL orange line) or Rabbit IgG isotype control (AC042, 2 μg/mL, blue line), followed by Alexa Fluor® 488 conjugated goat anti-rabbit pAb staining. Non-fluorescently stained cells were used as blank control (red line).
| Product name | DDDDK-Tag Rabbit mAb |
|---|---|
| Catalog No. | AE092 |
| Host species | Rabbit |
| Purification method | Affinity purification |
| Isotype | IgG |
| CloneNo. | ARC5111-01 |
| Immunogen | A synthetic peptide corresponding to DDDDK tag. |
|---|---|
| Sequence | DYKDDDDK |
| Gene ID | |
| Swiss Prot | |
| Synonyms | DDDDK; DDDDK tag; DDDDK-tag; DDDDK-Tag |
| Calculated MW | |
| Observed MW | 56kDa/50kDa/46kDa/68kDa |
| Reactivity | Species independent |
|---|---|
| Tested applications | WBIHC-PIF/ICCIPChIPChIP-seqRIPFCFC(Intra)ELISAMeDIPNucleotide ArrayDBFACSCoIPCUT&TagmeRIPInhibition |
| Recommended dilution |
|
| Storage buffer | Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3. |
| Key application | Western blotting Immunofluorescence Immunoprecipitation Flow Cytometry |
| Positive samples | 293F-GSK3B-Flag, 293F-BCAT2-Flag(N端), 293F-BCAT2-Flag(C端), 293F-COPB2-Flag-GFP(N端)293f-COPB2-Flag-GFP(C端) |
| Cellular location | |
| Customer validation | WB(Saccharomyces cerevisiae, Drosophila melanogaster, Mus musculus, Triticum aestivum, Homo sapiens, Danio rerio, Other, Oryza sativa, Arabidopsis thaliana, Chlorocebus aethiops) IF(Mus musculus) RIP(Homo sapiens) IP(Homo sapiens, Mus musculus) IP(Homo sapiens, Chlorocebus aethiops) Co-IP(Homo sapiens) ChIP(Mus musculus, Homo sapiens) Co-IP(Rattus norvegicus, Other, Arabidopsis thaliana) WB(Homo sapiens) FC(Homo sapiens) IHC(Homo sapiens, Chlorocebus aethiops) IF(Homo sapiens, Chlorocebus aethiops) |
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