• CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina (RK20265) from 10⁵ K562 cells with 1 Mu g Smad4 rabbit monoclonal antibody (STJ11101653) , along with a Goat Anti-rabbit IgG (H+L). The CUT&Tag results indicate the enrichment pattern of Smad4 in representative gene loci (JUNB) , as shown in figure.
  • Western blot analysis of extracts of various cell lines, using Smad4 antibody (STJ11101653) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-rabbit IgG (H+L) (STJS000856) at 1:10000 dilution. Lysates/proteins: 25 Mu g per lane. Blocking buffer: 3% non-fat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 10s.
  • Western blot analysis of extracts of various cell lines, using Smad4 antibody (STJ11101653) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-rabbit IgG (H+L) (STJS000856) at 1:10000 dilution. Lysates/proteins: 25 Mu g per lane. Blocking buffer: 3% non-fat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 3min.
  • Immunohistochemistry analysis of paraffin-embedded human liver cancer using [KO Validated] Smad4 rabbit monoclonal antibody (STJ11101653) at dilution of 1:100 (40x lens). Perform microwave antigen retrieval with 10 mM Tris/EDTA buffer pH 9. 0 before commencing with immunohistochemistry staining protocol.
  • Immunoprecipitation analysis of 300 Mu g extracts of 293T cells using 3 Mu g Smad4 antibody (STJ11101653). Western blot was performed from the immunoprecipitate using Smad4 antibody (STJ11101653) at a dilution of1:1000.
  • Chromatin immunoprecipitation analysis of extracts of HepG2 cells, using [KO Validated] Smad4 rabbit monoclonal antibody (STJ11101653) and rabbit IgG. The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.

Anti-SMAD4 antibody (320-552) [S3MR] (STJ11101653)

SKU:
STJ11101653

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Host: Rabbit
Applications: WB/IHC/IP
Reactivity: Human/Mouse/Rat
Note: STRICTLY FOR FURTHER SCIENTIFIC RESEARCH USE ONLY (RUO). MUST NOT TO BE USED IN DIAGNOSTIC OR THERAPEUTIC APPLICATIONS.
Short Description: Rabbit monoclonal antibody anti-Smad4 (320-552) is suitable for use in Western Blot, Immunohistochemistry and Immunoprecipitation research applications.
Clonality: Monoclonal
Clone ID: S3MR
Conjugation: Unconjugated
Isotype: IgG
Formulation: PBS with 0.01% Thimerosal, 0.05% BSA, 50% Glycerol, pH7.3.
Purification: Affinity purification
Dilution Range: WB 1:500-1:2000
IHC-P 1:50-1:200
IP 1:50-1:200
ChIP 1:50-1:200
CUT&Tag 10⁵ cells/1 Mu g
Storage Instruction: Store at-20°C for up to 1 year from the date of receipt, and avoid repeat freeze-thaw cycles.
Gene Symbol: SMAD4
Gene ID: 4089
Uniprot ID: SMAD4_HUMAN
Immunogen Region: 320-552
Immunogen: Recombinant fusion protein containing a sequence corresponding to amino acids 320-552 of human Smad4 (NP_005350.1).
Immunogen Sequence: PEYWCSIAYFEMDVQVGETF KVPSSCPIVTVDGYVDPSGG DRFCLGQLSNVHRTEAIERA RLHIGKGVQLECKGEGDVWV RCLSDHAVFVQSYYLDREAG RAPGDAVHKIYPSAYIKVFD LRQCHRQMQQQAATAQAAAA AQAAAVAGNIPGPGSVGGIA PAISLSAAAGIGVDDLRRLC ILRMSFVKGWGPDYPRQSIK ETPCWIEIHLHRALQLLDEV LHTMPIADPQPLD
Post Translational Modifications Phosphorylated by PDPK1. Monoubiquitinated on Lys-519 by E3 ubiquitin-protein ligase TRIM33. Monoubiquitination hampers its ability to form a stable complex with activated SMAD2/3 resulting in inhibition of TGF-beta/BMP signaling cascade. Deubiquitination by USP9X restores its competence to mediate TGF-beta signaling.
Function In muscle physiology, plays a central role in the balance between atrophy and hypertrophy. When recruited by MSTN, promotes atrophy response via phosphorylated SMAD2/4. MSTN decrease causes SMAD4 release and subsequent recruitment by the BMP pathway to promote hypertrophy via phosphorylated SMAD1/5/8. Acts synergistically with SMAD1 and YY1 in bone morphogenetic protein (BMP)-mediated cardiac-specific gene expression. Binds to SMAD binding elements (SBEs) (5'-GTCT/AGAC-3') within BMP response element (BMPRE) of cardiac activating regions. Common SMAD (co-SMAD) is the coactivator and mediator of signal transduction by TGF-beta (transforming growth factor). Component of the heterotrimeric SMAD2/SMAD3-SMAD4 complex that forms in the nucleus and is required for the TGF-mediated signaling. Promotes binding of the SMAD2/SMAD4/FAST-1 complex to DNA and provides an activation function required for SMAD1 or SMAD2 to stimulate transcription. Component of the multimeric SMAD3/SMAD4/JUN/FOS complex which forms at the AP1 promoter site.required for synergistic transcriptional activity in response to TGF-beta. May act as a tumor suppressor. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.
Protein Name Mothers Against Decapentaplegic Homolog 4
Mad Homolog 4
Mothers Against Dpp Homolog 4
Deletion Target In Pancreatic Carcinoma 4
Smad Family Member 4
Smad 4
Smad4
Hsmad4
Database Links Reactome: R-HSA-1181150
Reactome: R-HSA-1502540
Reactome: R-HSA-201451
Reactome: R-HSA-2173789
Reactome: R-HSA-2173795
Reactome: R-HSA-2173796
Reactome: R-HSA-3311021
Reactome: R-HSA-3315487
Reactome: R-HSA-452723
Reactome: R-HSA-5689880
Reactome: R-HSA-8941326
Reactome: R-HSA-8941855
Reactome: R-HSA-8952158
Reactome: R-HSA-9615017
Reactome: R-HSA-9617828
Reactome: R-HSA-9733709
Reactome: R-HSA-9735871
Reactome: R-HSA-9754189
Reactome: R-HSA-9823730
Cellular Localisation Cytoplasm
Nucleus
Cytoplasmic In The Absence Of Ligand
Migrates To The Nucleus When Complexed With R-Smad
Pdpk1 Prevents Its Nuclear Translocation In Response To Tgf-Beta
Alternative Antibody Names Anti-Mothers Against Decapentaplegic Homolog 4 antibody
Anti-Mad Homolog 4 antibody
Anti-Mothers Against Dpp Homolog 4 antibody
Anti-Deletion Target In Pancreatic Carcinoma 4 antibody
Anti-Smad Family Member 4 antibody
Anti-Smad 4 antibody
Anti-Smad4 antibody
Anti-Hsmad4 antibody
Anti-SMAD4 antibody
Anti-DPC4 antibody
Anti-MADH4 antibody

Information sourced from Uniprot.org

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