Revision 4

#12556Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-IC, FC-FP, ChIP, ChIP-seq

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

55-62

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q13950

Entrez-Gene Id:

860

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 5 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
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:6400 - 1:12800
Flow Cytometry (Fixed/Permeabilized) 1:1600 - 1:6400
Chromatin IP 1:100
Chromatin IP-seq 1:100

Storage

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 #68007.

Specificity / Sensitivity

RUNX2 (D1I7F) Rabbit mAb recognizes endogenous levels of total RUNX2 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala273 of human RUNX2 protein.

Background

Runt-related transcription factor 2 (RUNX2) is a member of the RUNX family of transcription factors. It is involved in osteoblast differentiation and skeletal morphogenesis. RUNX2 regulates the transcription of various genes, including osteopontin, bone sialoprotein, and osteocalcin, via binding to the core site of the enhancers or promoters (1-3). RUNX2 is crucial for the maturation of osteoblasts and both intramembranous and endochondral ossification. Mutations in the corresponding RUNX2 gene have been associated with the bone development disorder cleidocranial dysplasia (CCD) (4-6). RUNX2 is also abnormally expressed in various human cancers, including prostate and breast cancer. It plays an important role in migration, invasion, and bone metastasis of prostate and breast cancer cells (7-10).

  1. Viereck, V. et al. (2002) J Cell Biochem 86, 348-56.
  2. Willis, D.M. et al. (2002) J Biol Chem 277, 37280-91.
  3. Tu, Q. et al. (2008) J Cell Physiol 217, 40-7.
  4. Quack, I. et al. (1999) Am J Hum Genet 65, 1268-78.
  5. Cardoso, B.M. et al. (2010) Clin Dysmorphol 19, 150-2.
  6. Han, M.S. et al. (2010) J Cell Biochem 110, 97-103.
  7. Akech, J. et al. (2010) Oncogene 29, 811-21.
  8. van der Deen, M. et al. (2010) J Cell Biochem 109, 828-37.
  9. Barnes, G.L. et al. (2003) Cancer Res 63, 2631-7.
  10. Barnes, G.L. et al. (2004) Cancer Res 64, 4506-13.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 4
#12556

RUNX2 (D1L7F) Rabbit mAb

Western Blotting Image 1: RUNX2 (D1L7F) Rabbit mAb Expand Image
Western blot analysis of extracts from Saos-2 and LNCaP cells using RUNX2 (D1L7F) Rabbit mAb and β-Actin (D6A8) Rabbit mAb #8457.
Immunoprecipitation Image 1: RUNX2 (D1L7F) Rabbit mAb Expand Image
Immunoprecipitation of RUNX2 from Saos-2 cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or RUNX2 (D1L7F) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using RUNX2 (D1L7F) Rabbit mAb.
Immunofluorescence Image 1: RUNX2 (D1L7F) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of Saos-2 (left) and LNCaP (right) cells using RUNX2 (D1L7F) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Flow Cytometry Image 1: RUNX2 (D1L7F) Rabbit mAb Expand Image
Flow cytometric analysis of LNCap cells (blue, negative) and Saos-2 cells (green, positive) using RUNX2 (D1L7F) Rabbit mAb (solid lines) or a 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.
Chromatin Immunoprecipitation Image 1: RUNX2 (D1L7F) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Saos-2 cells and RUNX2 (D1L7F) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across RXRG gene.
Chromatin Immunoprecipitation Image 2: RUNX2 (D1L7F) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Saos-2 cells and RUNX2 (D1L7F) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 1 (upper), including RXRG gene (lower).
Chromatin Immunoprecipitation Image 3: RUNX2 (D1L7F) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Saos-2 cells and either RUNX2 (D1L7F) 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 RUNX1 Intron 1 primers, SimpleChIP® Human RUNX2 Promoter Primers #12376, human FRA10AC1 intron 1 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.