全部商品分类
用小程序，查商品更便捷
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys460 of human HIF-1α protein.
Product Usage Information
For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Simple Western™ | 1:50 - 1:250 |
| Chromatin IP | 1:50 |
| Chromatin IP-seq | 1:50 |
| CUT&RUN | 1:50 |
Specificity/Sensitivity
Species Reactivity:
Human, Mouse, Rat, Monkey
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
参考图片
CUT&RUN was performed with MCF7 cells treated with cobalt chloride (100 μM) overnight and HIF-1α (D2U3T) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ARRDC3 Downstream Primers #75671 and SimpleChIP® Human α Satelite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Western blot analysis of extracts from various cell lines, untreated (-) or treated with either DMOG (1 mM, 6 hr; +) or cobalt chloride (100 µM, 4 hr; +) as indicated, using HIF-1α (D2U3T) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Simple Western™ analysis of lysates (1.0 mg/mL) from Raji cells using HIF-1α (D2U3T) Rabbit mAb #14179. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with cobalt chloride (100 μM) overnight and HIF-1α (D2U3T) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across the PCAT6 gene.
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with cobalt chloride (100 μM) overnight and HIF-1α (D2U3T) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 1 (upper), including the PCAT6 gene (lower).
Chromatin immunopreciptations were performed with cross-linked chromatin from MCF7 cells treated with cobalt chloride (100 μM) overnight and either HIF-1α (D2U3T) Rabbit mAb or Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ARRDC3 Downstream Primers #75671, human ERRFI1 upstream primers, and SimpleChIP® Human α Satelite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT&RUN was performed with MCF7 cells treated with cobalt chloride (100 μM) overnight and HIF-1α (D2U3T) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across HIF-1α gene.
CUT&RUN was performed with MCF7 cells treated with cobalt chloride (100 μM) overnight and HIF-1α (D2U3T) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 14 (upper), including HIF-1α gene (lower).