Phospho-RAF1 (Ser259) Recombinant monoclonal antibody

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  WB analysis of HeLa using 88430-1-RR

  Non-treated HeLa cells, phosphatase inhibitor treated HeLa cells and λ phosphatase treated HeLa cells were subjected to SDS PAGE followed by western blot with 88430-1-RR (Phospho-RAF1 (Ser259) antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Alpha Tubulin (HRP-80762) antibody as a loading control.

Phospho-RAF1 (Ser259) Uni-rAb^® Recombinant Antibody for WB, ELISA

Cat No. 88430-1-RR

Clone No.250953G5

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, ELISA

Synonyms

c-Raf (Ser259), p-c-Raf, p-c-Raf (Ser259), Phospho-c-Raf, Phospho-c-Raf (Ser259)

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Tested Applications

• Positive WB detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

88430-1-RR targets Phospho-RAF1 (Ser259) in WB, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen:

  Peptide

• Full Name: v-raf-1 murine leukemia viral oncogene homolog 1
• Calculated Molecular Weight: 648 aa, 73 kDa
• Observed Molecular Weight: 70-75 kDa
• GenBank Accession Number: BC018119
• Gene Symbol: RAF1
• Gene ID (NCBI): 5894
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: P04049
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Raf-1 proto-oncogene, serine/threonine kinase(RAF1), is a MAP kinase kinase kinase (MAP3K), which functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Phospho-RAF1 (Ser259) is a key inhibitory modification of CRAF within the RAS-MEK-ERK cascade. Phosphorylated mainly by PKA and Akt, this residue recruits 14-3-3 proteins to lock RAF1 in an autoinhibited cytoplasmic conformation, blocking its membrane recruitment and binding to GTP-Ras. Dephosphorylation by PP2A relieves suppression to trigger ERK-driven proliferation. Elevated p-RAF1 (S259) dampens oncogenic MAPK signaling in tumors, while loss of this phosphorylation causes sustained pathway hyperactivation linked to Noonan syndrome and multiple malignancies. (PMID: 11971957, 11756411)