Revision 4

#12556

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, 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 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.

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).

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

W: 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

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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 , (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

Cell Signaling Technology Logo
Western blot analysis of extracts from Saos-2 and LNCaP cells using RUNX2 (D1L7F) Rabbit mAb and β-Actin (D6A8) Rabbit mAb #8457.
Western Blotting Image 1: RUNX2 (D1L7F) Rabbit mAb
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.
Immunoprecipitation Image 1: RUNX2 (D1L7F) Rabbit mAb
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).
Immunofluorescence Image 1: RUNX2 (D1L7F) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

Cell Signaling Technology Logo
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.
Flow Cytometry Image 1: RUNX2 (D1L7F) Rabbit mAb
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 1: RUNX2 (D1L7F) Rabbit mAb
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 2: RUNX2 (D1L7F) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

Cell Signaling Technology Logo
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.
Chromatin Immunoprecipitation Image 3: RUNX2 (D1L7F) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.