Cell Signaling Technology

Product Pathways - Transcription Factors

Helios (D8W4X) XP® Rabbit mAb #42427

No. Size Price
42427S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
42427 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse, Endogenous 58, 63-70, 85 Rabbit IgG
IP 1:200
F 1:100
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, ChIP=Chromatin IP,

Specificity / Sensitivity

Helios (D8W4X) XP® Rabbit mAb recognizes endogenous levels of total Helios protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly73 of human Helios protein.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of RL-7 (blue) and Jurkat (green) cells using Helios (D8W4X) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary anitbody.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Jurkat cells and either 10 μl of Helios (D8W4X) XP® Rabbit mAb or 2 μl of 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 IKZF3 Promoter Primers #99663, human IL-2 promoter primers, 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 Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Helios (D8W4X) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

IP

IP

Immunoprecipitation of Helios from Jurkat cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Helios (D8W4X) XP® Rabbit mAb. Western blot analysis was performed using Helios (D8W4X) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, untransfected (-) or transfected with a construct expressing Myc-tagged full-length human Helios (hHelios-Myc; +), using Helios (D8W4X) XP® Rabbit mAb.

Background

Helios (Ikaros family zinc finger 2, IZKF2) is an Ikaros family transcription factor composed of several zinc fingers that mediate DNA binding and homodimerization or heterodimerization with other Ikaros family proteins (1,2). In the hematopoietic system, Helios expression is restricted to T cells and early hematopoietic progenitors (1,2). In regulatory T cells, expression of Helios contributes to an anergic phenotype by binding to the IL-2 promoter and suppressing IL-2 transcription (3). In addition, alteration of the corresponding Helios gene IZKF2 is one hallmark of low-hypodiploid acute lymphoblastic leukemia (4).

  1. Hahm, K. et al. (1998) Genes Dev 12, 782-96.
  2. Kelley, C.M. et al. (1998) Curr Biol 8, 508-15.
  3. Baine, I. et al. (2013) J Immunol 190, 1008-16.
  4. Holmfeldt, L. et al. (2013) Nat Genet 45, 242-52.

Application References

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Protocols

Companion Products


For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

Tween is a registered trademark of ICI Americas, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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