Revision 7

#73885

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, IHC-Bond, IHC-P, ELISA, IF-IC, FC-FP

Reactivity:
H M R Mk

Sensitivity:
Endogenous

MW (kDa):
70

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q13485

Entrez-Gene Id:
4089

Product Usage Information

This product is the carrier free version of product #46535. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN, or CUT&Tag assays. If you require a carrier-free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

BSA and Azide Free antibodies are quality control tested by size exclusion chromatography (SEC) to determine antibody integrity.

Formulation

Supplied in 1X PBS (10 mM Na2HPO4, 3 mM KCl, 2 mM KH2PO4, and 140 mM NaCl (pH 7.8)). BSA and Azide Free.

For standard formulation of this product see product #46535.

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity/Sensitivity

SMAD4 (D3R4N) XP® Rabbit mAb (BSA and Azide Free) recognizes endogenous levels of total SMAD4 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein corresponding to human SMAD4 protein. The epitope has been mapped to residues surrounding Ser315.

Background

Members of the SMAD family of signal transduction molecules are components of a critical intracellular pathway that transmits TGF-β signals from the cell surface into the nucleus. Three distinct classes of SMADs have been defined: the receptor-regulated SMADs (R-SMADs), which include SMAD1, 2, 3, 5, 9; the common-mediator SMAD (co-SMAD), SMAD4; and the antagonistic or inhibitory SMADs (I-SMADs), SMAD6 and 7 (1-5). Activated type I receptors associate with specific R-SMADs and phosphorylate them on a conserved SSXS motif in the carboxy-terminus. Phosphorylated R-SMADS dissociate from the receptor and form a heteromeric complex with SMAD4, initiating translocation of the heteromeric SMAD complex to the nucleus. Once in the nucleus, SMADs recruit a variety of DNA binding proteins that function to regulate transcriptional activity (6-8).

Species Reactivity

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

Applications Key

W: Western Blotting IHC-Bond: IHC Leica Bond IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

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 G: Goat All: All Species Expected

Trademarks and Patents

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

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

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

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

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 7

CST Logo
Western blot analysis of extracts from various cell lines using SMAD4 (D3R4N) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Specificity of the antibody is confirmed by the absence of signal in extracts from HT-29 and COLO 205 cells, both of which contain Smad4 null mutations. Data were generated using the standard formulation of this product.
Western Blotting Image 1: SMAD4 (D3R4N) XP<sup>®</sup> Rabbit mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human neuroendocrine lung carcinoma using SMAD4 (D3R4N) XP® Rabbit mAb performed on the Leica® BOND™ Rx. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 1: SMAD4 (D3R4N) XP<sup>®</sup> Rabbit mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human infiltrating papillary carcinoma of the breast using SMAD4 (D3R4N) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 1: SMAD4 (D3R4N) XP<sup>®</sup> Rabbit mAb (BSA and Azide Free)
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 7

CST Logo
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using SMAD4 (D3R4N) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 2: SMAD4 (D3R4N) XP<sup>®</sup> Rabbit mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human ductal carcinoma of the breast using SMAD4 (D3R4N) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 3: SMAD4 (D3R4N) XP<sup>®</sup> Rabbit mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded COLO 320 cell pellet (left, positive) or COLO 205 cell pellet (right, negative) using SMAD4 (D3R4N) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 4: SMAD4 (D3R4N) XP<sup>®</sup> Rabbit mAb (BSA and Azide Free)
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 7

CST Logo
Confocal immunofluorescent analysis of HaCaT cells (positive, top panel) or HT-29 cells (negative, bottom panel), either serum-starved (left panel) or serum-starved and treated with Human Transforming Growth Factor β1 (hTGF-β1) #8915 (7 ng/mL, 1 hr; right panel), using Smad4 (D3R4N) XP® Rabbit mAb (green) and DyLight™ 554 Phalloidin #13054 (red). Data were generated using the standard formulation of this product.
Immunofluorescence Image 1: SMAD4 (D3R4N) XP<sup>®</sup> Rabbit mAb (BSA and Azide Free)
Flow cytometric analysis of HT-29 cells (blue) and HCT 116 cells (green) using SMAD4 (D3R4N) XP® 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. Data were generated using the standard formulation of this product.
Flow Cytometry Image 1: SMAD4 (D3R4N) XP<sup>®</sup> Rabbit mAb (BSA and Azide Free)
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.