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AB279903

Phospho-Raf1 (S301) ELISA Kit

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Phospho-Raf1 (S301) ELISA Kit is a Semi-quantitative ELISA for the measurement of Phospho-Raf1 (S301) in Human in Cell/Tissue Extracts samples.

View Alternative Names

RAF, RAF1, RAF proto-oncogene serine/threonine-protein kinase, Proto-oncogene c-RAF, Raf-1, cRaf

3 Images
Sandwich ELISA - Phospho-Raf1 (S301) ELISA Kit (AB279903)
  • sELISA

Supplier Data

Sandwich ELISA - Phospho-Raf1 (S301) ELISA Kit (AB279903)

Positive Control.

Jurkat cells were treated with PMA.

Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer.

Serial dilutions of lysates were analyzed in this ELISA.

Sandwich ELISA - Phospho-Raf1 (S301) ELISA Kit (AB279903)
  • sELISA

Supplier Data

Sandwich ELISA - Phospho-Raf1 (S301) ELISA Kit (AB279903)

Jurkat cells treated with/without PMA. Jurkat cells were untreated or treated with PMA.

Sandwich ELISA - Phospho-Raf1 (S301) ELISA Kit (AB279903)
  • sELISA

Supplier Data

Sandwich ELISA - Phospho-Raf1 (S301) ELISA Kit (AB279903)

Jurkat cells treated with/without PMA. Jurkat cells were untreated or treated with PMA.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Reacts with

Human

Assay type

Semi-quantitative

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "ELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

Phospho-Raf1 (S301) ELISA Kit (ab279903) 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 Raf1 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-Raf1. An anti-pan Raf1 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Raf1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-phospho-Raf1 (S301) antibody is used to detect phosphorylated Raf1. After washing away unbound antibody, HRP conjugated anti-rabbit IgG 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 Raf1 (S301) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

REACH authorisation
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.

What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab279903-1x96Tests", "assetComponentDetails": [ { "size":"1 x 5 mL", "name":"2X Cell lysate buffer", "number":"AB279903-CMP09", "productcode":"" }, { "size":"1 x 25 mL", "name":"20X Wash Buffer", "number":"AB279903-CMP03", "productcode":"" }, { "size":"1 x 8 mL", "name":"Stop Solution", "number":"AB279903-CMP07", "productcode":"" }, { "size":"1 x 1 Vial", "name":"Positive Control - Jurkat cell lysate", "number":"AB279903-CMP01", "productcode":"" }, { "size":"1 x 15 mL", "name":"5X Assay Diluent", "number":"AB279903-CMP05", "productcode":"" }, { "size":"1 x 12 mL", "name":"TMB One-Step Substrate Reagent", "number":"AB279903-CMP08", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Pan-Raf1 Coated Microplate", "number":"AB279903-CMP04", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Rabbit anti-phospho-Raf1 (S301)-antibody", "number":"AB279903-CMP06", "productcode":"" }, { "size":"1 x 25 µL", "name":"HRP-conjugated anti-rabbit IgG concentrate (500X)", "number":"AB279903-CMP02", "productcode":"" } ] } } }

Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Raf1 also known as c-Raf or Raf-1 is a serine/threonine-protein kinase that plays an important role in cell division differentiation and survival. Its molecular weight is approximately 74 kDa. Raf1 is part of the MAPK/ERK signaling pathway and is ubiquitously expressed in many tissues including the brain heart and liver. As a component of the signaling cascade its activation translates extracellular signals into intracellular responses.
Biological function summary

Raf1 regulates important cellular processes by activating downstream kinases in response to external stimuli. Raf1 forms a complex with other proteins such as Ras facilitating its role as an essential component of the MAPK/ERK pathway. Upon activation by Ras Raf1 phosphorylates and activates MEK1 and MEK2 which in turn activate the ERK1 and ERK2. This signaling axis is involved in controlling gene expression and cellular proliferation.

Pathways

Raf1 is integrally involved in the MAPK/ERK pathway which is critical for transducing signals from growth factors and mitogens. It relates closely with proteins such as Ras MEK and ERK in this pathway. The pathway is important for regulating cellular responses to various stimuli and is particularly involved in processes such as cell cycle control and apoptosis.

Raf1 has associations with certain types of cancer and Noonan syndrome. In various cancers mutations or dysregulation of Raf1 or related components such as Ras can lead to uncontrolled cellular proliferation. In Noonan syndrome Raf1 mutations result in anomalies in the Ras/MAPK pathway which can impact normal development. Raf1’s interactions in these pathways indicate its relevance as a potential target for therapeutic intervention.

Product protocols

Target data

Serine/threonine-protein kinase that acts as a regulatory link between the membrane-associated Ras GTPases and the MAPK/ERK cascade, and this critical regulatory link functions as a switch determining cell fate decisions including proliferation, differentiation, apoptosis, survival and oncogenic transformation. RAF1 activation initiates a mitogen-activated protein kinase (MAPK) cascade that comprises a sequential phosphorylation of the dual-specific MAPK kinases (MAP2K1/MEK1 and MAP2K2/MEK2) and the extracellular signal-regulated kinases (MAPK3/ERK1 and MAPK1/ERK2). The phosphorylated form of RAF1 (on residues Ser-338 and Ser-339, by PAK1) phosphorylates BAD/Bcl2-antagonist of cell death at 'Ser-75'. Phosphorylates adenylyl cyclases : ADCY2, ADCY5 and ADCY6, resulting in their activation. Phosphorylates PPP1R12A resulting in inhibition of the phosphatase activity. Phosphorylates TNNT2/cardiac muscle troponin T. Can promote NF-kB activation and inhibit signal transducers involved in motility (ROCK2), apoptosis (MAP3K5/ASK1 and STK3/MST2), proliferation and angiogenesis (RB1). Can protect cells from apoptosis also by translocating to the mitochondria where it binds BCL2 and displaces BAD/Bcl2-antagonist of cell death. Regulates Rho signaling and migration, and is required for normal wound healing. Plays a role in the oncogenic transformation of epithelial cells via repression of the TJ protein, occludin (OCLN) by inducing the up-regulation of a transcriptional repressor SNAI2/SLUG, which induces down-regulation of OCLN. Restricts caspase activation in response to selected stimuli, notably Fas stimulation, pathogen-mediated macrophage apoptosis, and erythroid differentiation.
See full target information RAF1 phospho S301
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