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Monoclonal antibody is produced by immunizing animals with a synthetic peptide containing the influenza hemagglutinin epitope (YPYDVPDYA).
Product Usage Information
For optimal ChIP 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.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Simple Western™ | 1:50 - 1:250 |
| Immunoprecipitation | 1:50 |
| Immunohistochemistry (Paraffin) | 1:800 - 1:3200 |
| Immunofluorescence (Immunocytochemistry) | 1:800 - 1:1600 |
| Flow Cytometry (Fixed/Permeabilized) | 1:800 - 1:1600 |
| Chromatin IP | 1:50 |
Specificity/Sensitivity
Species Reactivity:
All Species Expected
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.
For a carrier free (BSA and azide free) version of this product see product #86124.
参考图片
293T cells were either untransfected (left panel) or transfected with an HA-tagged human CBP construct (right panel), then treated with Forskolin #3828 (30 µM) . Chromatin immunoprecipitations were performed with cross-linked chromatin from cells and HA-Tag (C29F4) Rabbit mAb, CBP (D9B6) 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 human ALS2 exon 1 primers, SimpleChIP® Human NR4A3 Promoter Primers #4829, and SimpleChIP® Human α Satellite 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 HeLa cells, untransfected or transfected with either HA-FoxO4 or HA-Akt3, using HA-Tag (C29F4) Rabbit mAb.
Simple Western™ analysis of lysates (0.1 mg/mL) from COS-7 cells, transfected with a construct expressing HA-tagged Stat3 using HA-Tag (C29F4) Rabbit mAb #3724. The virtual lane view (left) shows a single 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.
Immunoprecipitation of HA-FoxO4 tag protein from 293T transfected cell extracts. Lane 1 is 10% input, lane 2 is HA-Tag (C29F4) Rabbit mAb, and lane 3 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. Western blot was performed using HA-Tag (C29F4) Rabbit mAb. A conformation specific secondary antibody was used to avoid reactivity with IgG.
Immunohistochemical analysis of paraffin-embedded COS cells, HA-Tag transfected (left) or untransfected (right), using HA-Tag (C29F4) Rabbit mAb.
Confocal immunofluorescent analysis of COS cells, transfected with an HA-tagged protein (left) or mock-transfected (right), using HA-Tag (C29F4) Rabbit mAb (green). Actin filaments have been labeled using DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
Flow cytometric analysis of 293T cells untransfected (blue) or transfected with HA-tagged Akt (green), using HA-Tag (C29F4) Rabbit mAb (solid lines) or concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines).Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.