Skip to main content

Phospho-Smad4 (T277) and Total Smad4 ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-Smad4 (T277) and Total Smad4 in Human in Cell Lysate samples.

Be the first to review this product! Submit a review

Images

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Assay type

Semi-quantitative

Reactive species

Human

Loading...
Loading...
Loading...
Loading...
Loading...
Loading...
Loading...
Loading...

Reactivity data

Application

ELISA

Reactivity

Reacts

Dilution info

-

Notes

-

What's included?

1 x 96 Tests
Components
20X Wash Buffer
1 x 25 mL
2X Cell lysate buffer
1 x 5 mL
300X Streptavidin-Conjugated HRP
1 x 200 µL
5X Assay Diluent
1 x 15 mL
Biotinylated anti-pan-Smad4 detection antibody
1 x 1 Vial
HRP-conjugated anti-rabbit IgG concentrate (1000X)
1 x 25 µL
Pan-Smad4 Coated Microplate
1 x 1 Unit
Positive Control - treated Jurkat cell lysate
1 x 1 Vial
Rabbit anti-phospho-Smad4 (T277)-antibody
1 x 1 Vial
Stop Solution
1 x 8 mL
TMB One-Step Substrate Reagent
1 x 12 mL

Recommended products

  1. Loading...
  2. Loading...
  3. Loading...
  4. Loading...

Phospho-Smad4 (T277) and Total Smad4 ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-Smad4 (T277) and Total Smad4 in Human in Cell Lysate samples.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Assay type

Semi-quantitative

Reactive species

Human

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

Phospho-Smad4 (T277) and Total Smad4 ELISA Kit (ab279932) is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated Smad4 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-Smad4 and total Smad4. An anti-pan Smad4 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Smad4 present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho Smad4 (T277) antibody is added to detect phosphorylated Smad4. In the remaining wells, biotinylated anti-pan-Smad4 antibody is used to detect pan Smad4. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-Streptavidin is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Smad4 (T277) or pan Smad4 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Supplementary info

Biological function summary

Smad4 influences cell proliferation differentiation and apoptosis by mediating signals from TGF-beta cytokines. It is part of the Smad protein family acting as a transcriptional controller. Upon TGF-beta receptor activation Smad4 forms complexes with Smad2 and Smad3 translocating to the nucleus to regulate genes imperative for cellular homeostasis. Its role in cell cycle regulation underlines its contribution to normal cellular functions and its potential involvement in disorders.

Activity summary

Smad4 also known as DPC4 or MADH4 is a central protein in the TGF-beta signaling pathway with a molecular mass of approximately 60 kDa. It acts as a signal transducer that facilitates communication from the cell surface to the nucleus. Smad4 is broadly expressed in various tissues playing an important role in the regulation of cellular processes. It forms a complex with receptor-regulated Smads (R-Smads) to translocate to the nucleus where it influences gene transcription.

Pathways

Smad4 operates within the TGF-beta pathway linking extracellular signals to nuclear transcription alterations. It participates in the regulation of epithelial-mesenchymal transition (EMT) a process important for development and tumor progression. In these pathways Smad4 interacts closely with Smad2 and Smad3 orchestrating various cellular responses to external stimuli through transcriptional management.

Associated diseases and disorders

Smad4 is highly related to cancer and juvenile polyposis syndrome. Mutations or deletions in Smad4 disrupt its function contributing to the progression of pancreatic cancer and colorectal cancer among others. Within these contexts Smad4 connects strongly to other proteins like p21 and cyclin-dependent kinase inhibitors which are important in cell cycle arrest and impede tumor growth.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

2 product images

  • Sandwich ELISA - Phospho-Smad4 (T277) and Total Smad4 ELISA Kit (ab279932), expandable thumbnail

    Sandwich ELISA - Phospho-Smad4 (T277) and Total Smad4 ELISA Kit (ab279932)

    Positive Control.

    Jurkat cells were treated with PV (Pervanadate).

    Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer.

    Serial dilutions of lysates were analyzed in this ELISA.

  • Sandwich ELISA - Phospho-Smad4 (T277) and Total Smad4 ELISA Kit (ab279932), expandable thumbnail

    Sandwich ELISA - Phospho-Smad4 (T277) and Total Smad4 ELISA Kit (ab279932)

    Jurkat cells treated with/without CPT.

    Jurkat cells were untreated or treated with CPT. Cell lysates were analyzed using this phosphoELISA and Western Blot.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

There was a problem

We can’t download that datasheet. Please try again. If you need help, contact our Customer Services team at technical@abcam.com