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  1. Link

    products/primary-antibodies/smad4-antibody-ep618y-ab40759.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Smad4 antibody [EP618Y] (ab40759)

  • Datasheet
  • SDS
Reviews (9) Submit a question References (136)

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Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)
  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)
  • Anti-Smad4 antibody [EP618Y] (ab40759)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP618Y] to Smad4
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free HRP

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Overview

  • Product name

    Anti-Smad4 antibody [EP618Y]
    See all Smad4 primary antibodies
  • Description

    Rabbit monoclonal [EP618Y] to Smad4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Smad4 aa 500 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: Q13485
    (Peptide available as ab228416)

  • Positive control

    • WB: Wild type HAP1 whole cell lysate; HepG2, Jurkat, NIH/3T3, PC-12, Ramos, C6 and SH-SY5Y cell lysates; Mouse embryo, skin and lung tissue lysates; Human skin, lung and artery tissue lysates. IHC-P: Human lung carcinoma and breast carcinoma tissues.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP618Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • SMADs
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • SMADs
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Cytoplasmic
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • Other
    • Cardiovascular
    • Heart
    • Apoptosis
    • Cardiovascular
    • Heart
    • Cardiogenesis
    • Transcription factors/regulators
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Transcription factors
    • Cardiovascular
    • Vasculature
    • Endothelium
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia

Associated products

  • Alternative Versions

    • HRP Anti-Smad4 antibody [EP618Y] (ab195554)
    • Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human Smad4 protein (ab81764)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Ramos whole cell lysate (ab3955)
    • NIH/3T3 whole cell lysate (ab7179)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab40759 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB (3)
1/5000. Detects a band of approximately 60 kDa (predicted molecular weight: 65 kDa).
IHC-P (4)
1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Notes
WB
1/5000. Detects a band of approximately 60 kDa (predicted molecular weight: 65 kDa).
IHC-P
1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Application notes
Is unsuitable for Flow Cyt,ICC/IF or IP.

Target

  • Function

    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 syngernistic transcriptional activity in response to TGF-beta. May act as a tumor suppressor.
  • Involvement in disease

    Defects in SMAD4 are a cause of pancreatic cancer (PNCA) [MIM:260350].
    Defects in SMAD4 are a cause of juvenile polyposis syndrome (JPS) [MIM:174900]; also known as juvenile intestinal polyposis (JIP). JPS is an autosomal dominant gastrointestinal hamartomatous polyposis syndrome in which patients are at risk for developing gastrointestinal cancers. The lesions are typified by a smooth histological appearance, predominant stroma, cystic spaces and lack of a smooth muscle core. Multiple juvenile polyps usually occur in a number of Mendelian disorders. Sometimes, these polyps occur without associated features as in JPS; here, polyps tend to occur in the large bowel and are associated with an increased risk of colon and other gastrointestinal cancers.
    Defects in SMAD4 are a cause of juvenile polyposis/hereditary hemorrhagic telangiectasia syndrome (JP/HHT) [MIM:175050]. JP/HHT syndrome phenotype consists of the coexistence of juvenile polyposis (JIP) and hereditary hemorrhagic telangiectasia (HHT) [MIM:187300] in a single individual. JIP and HHT are autosomal dominant disorders with distinct and non-overlapping clinical features. The former, an inherited gastrointestinal malignancy predisposition, is caused by mutations in SMAD4 or BMPR1A, and the latter is a vascular malformation disorder caused by mutations in ENG or ACVRL1. All four genes encode proteins involved in the transforming-growth-factor-signaling pathway. Although there are reports of patients and families with phenotypes of both disorders combined, the genetic etiology of this association is unknown.
    Defects in SMAD4 may be a cause of colorectal cancer (CRC) [MIM:114500].
  • Sequence similarities

    Belongs to the dwarfin/SMAD family.
    Contains 1 MH1 (MAD homology 1) domain.
    Contains 1 MH2 (MAD homology 2) domain.
  • Domain

    The MH1 domain is required for DNA binding.
    The MH2 domain is required for both homomeric and heteromeric interactions and for transcriptional regulation. Sufficient for nuclear import.
  • Post-translational
    modifications

    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. Deubiqitination by USP9X restores its competence to mediate TGF-beta signaling.
  • Cellular localization

    Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with R-SMAD.
  • Target information above from: UniProt accession Q13485 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4089 Human
    • Entrez Gene: 17128 Mouse
    • Entrez Gene: 50554 Rat
    • Omim: 600993 Human
    • SwissProt: Q13485 Human
    • SwissProt: P97471 Mouse
    • SwissProt: O70437 Rat
    • Unigene: 75862 Human
    • Unigene: 100399 Mouse
    • Unigene: 9774 Rat
    see all
  • Alternative names

    • (Small) Mothers Against Decapentaplegic antibody
    • Deleted in Pancreatic Carcinoma 4 antibody
    • Deleted in Pancreatic Carcinoma antibody
    • Deleted in pancreatic carcinoma locus 4 antibody
    • Deletion target in pancreatic carcinoma 4 antibody
    • DPC 4 antibody
    • DPC4 antibody
    • hSMAD4 antibody
    • JIP antibody
    • MAD homolog 4 antibody
    • MAD mothers against decapentaplegic Drosophila homolog 4 antibody
    • MAD mothers against decapentaplegic homolog 4 antibody
    • MADH 4 antibody
    • MADH4 antibody
    • Med antibody
    • Medea antibody
    • Mothers against decapentaplegic homolog 4 antibody
    • Mothers against decapentaplegic, Drosophila, homolog of, 4 antibody
    • Mothers against DPP homolog 4 antibody
    • MYHRS antibody
    • OTTHUMP00000163548 antibody
    • SMA- and MAD-related protein 4 antibody
    • SMAD 4 antibody
    • SMAD family member 4 antibody
    • SMAD mothers against DPP homolog 4 antibody
    • SMAD4 antibody
    • SMAD4_HUMAN antibody
    see all

Images

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: SMAD4 knockout  HAP1 whole cell lysate (20 µg)
    Lane 3: HepG2 whole cell lysate (20 µg)
    Lane 4: Jurkat whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab40759 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab40759 was shown to specifically react with SMAD4 in wild type HAP1 cells. No band was observed when SMAD4 knockout HAP1 samples were used. Wild-type and SMAD4 knockout samples were subjected to SDS-PAGE. Ab40759 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    All lanes : Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution (purified)

    Lane 1 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) cell lysate
    Lane 2 : Ramos (Human Burkitt's lymphoma cell line) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 65 kDa
    Observed band size: 60 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Smad4 with purified ab40759 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    All lanes : Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution

    Lane 1 : NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate
    Lane 2 : Mouse embryo tissue lysate
    Lane 3 : Mouse skin tissue lysate
    Lane 4 : Mouse lung tissue lysate
    Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate
    Lane 6 : C6 cell lysate
    Lane 7 : Rat skin tissue lysate
    Lane 8 : Rat lung tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 65 kDa
    Observed band size: 60 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    All lanes : Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution

    Lane 1 : SW480 (Human colorectal adenocarcinoma cell line) cell lysate
    Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate
    Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate
    Lane 4 : Human skin tissue lysate
    Lane 5 : Human lung tissue lysate
    Lane 6 : Human artery tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 65 kDa
    Observed band size: 60 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Different batches of ab40759 were tested on Ramos (Human Burkitt's lymphoma B lymphocyte) lysate at 0.7 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 60 kDa.
  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Anti-Smad4 antibody [EP618Y] (ab40759) at 1/10000 dilution (purified) + NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 65 kDa
    Observed band size: 60 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution (purified) + PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 65 kDa
    Observed band size: 60 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)
    Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution (unpurified) + SHSY5Y (Human neuroblastoma cell line from bone marrow) cell lysate at 10 µg

    Predicted band size: 65 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling Smad4 with unpurified ab40759 at a 1/100 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)This image is courtesy of an anonymous Abreview

    Unpurified ab40759 staining Smad4 in rat femur tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 20 minutes at 22°C; antigen retrieval was by heat mediation in a citrate buffer pH6.0. Samples were incubated with primary antibody (1/200 in blocking buffer) for 2 hours at 20°C. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

    See Abreview

  • Anti-Smad4 antibody [EP618Y] (ab40759)
    Anti-Smad4 antibody [EP618Y] (ab40759)

Protocols

  • Abreview protocol for ChIP using ab40759
  • Recommended FACS protocol

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (136)

Publishing research using ab40759? Please let us know so that we can cite the reference in this datasheet.

ab40759 has been referenced in 136 publications.

  • Long C  et al. FOXO3 is targeted by miR-223-3p and promotes osteogenic differentiation of bone marrow mesenchymal stem cells by enhancing autophagy. Hum Cell 34:14-27 (2021). PubMed: 32920731
  • Zhao S  et al. Effects of Smad4 on the expression of caspase-3 and Bcl-2 in human gingival fibroblasts cultured on 3D PLGA scaffolds induced by compressive force. Int J Mol Med 47:N/A (2021). PubMed: 33495811
  • Zhang T  et al. Proteomics reveals the function reverse of MPSSS-treated prostate cancer-associated fibroblasts to suppress PC-3 cell viability via the FoxO pathway. Cancer Med 10:2509-2522 (2021). PubMed: 33704935
  • Wang W  et al. Positive feedback regulation between USP15 and ERK2 inhibits osteoarthritis progression through TGF-ß/SMAD2 signaling. Arthritis Res Ther 23:84 (2021). PubMed: 33726807
  • Xu R  et al. MicroRNA-181a promotes epithelial-mesenchymal transition in esophageal squamous cell carcinoma via the TGF-ß/Smad pathway. Mol Med Rep 23:N/A (2021). PubMed: 33760133
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-9 of 9 Abreviews or Q&A

Western blot abreview for Anti-Smad4 antibody [EP618Y]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Coturnix japonica Tissue lysate - whole (yolk sac membrane from embryonic day 3 embryos)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10% gel)
Loading amount
50 µg
Specification
yolk sac membrane from embryonic day 3 embryos
Blocking step
Gelatin buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.25% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Miss. HAN-JEN LIN

Verified customer

Submitted Apr 17 2018

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad4 antibody [EP618Y]

Average
Abreviews
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako
Permeabilization
No
Specification
Liver
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 01 2016

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad4 antibody [EP618Y]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (postnatal day 21 mouse ovary)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.01M citric acide/sodium citrate
Permeabilization
No
Specification
postnatal day 21 mouse ovary
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
4% PFA
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 05 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-Smad4 antibody [EP618Y]

Good
Abreviews
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (A549)
Permeabilization
Yes - 0.25% Triton X-100/PBS
Specification
A549
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 11 2013

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad4 antibody [EP618Y]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Lung tissue)
Specification
Lung tissue
Fixative
HOPE
Antigen retrieval step
None
Permeabilization
No
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 27 2013

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad4 antibody [EP618Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (femur)
Specification
femur
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrated buffer, pH6.0
Permeabilization
No
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 23 2012

Western blot abreview for Anti-Smad4 antibody [EP618Y]

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (lymphoblats)
Loading amount
40 µg
Specification
lymphoblats
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris Gel, MOPS Buffer)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Lukas Rambousek

Verified customer

Submitted Dec 15 2010

ChIP abreview for Anti-Smad4 antibody [EP618Y]

Average
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - whole cell (MDA-MB-231 Cells treated with 2 ng/µl TGF-beta1)
Negative control
Polyclonal anti-HA antibody (for detection of non-specific binding). The graph is shown as a "fold enrichment" relative to the average detected HA binding.
Specification
MDA-MB-231 Cells treated with 2 ng/µl TGF-beta1
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 14 2007

Western blot abreview for Anti-Smad4 antibody [EP618Y]

Good
Abreviews
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (colon cancer)
Loading amount
50 µg
Specification
colon cancer
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 11 2006

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