Collaborator ELK Biotechnology
Primary AntibodyISO 9001:2015
ABM99312

Raf-1 (phospho Ser296) rabbit pAb

ISO 9001|ABMIUM Verified™|Antibody

Raf-1 (phospho Ser296) rabbit pAb from ELK Biotechnology Read more ›

Raf-1 (phospho Ser296) rabbit pAb
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Raf-1 (phospho Ser296) rabbit pAb

Product Description

Raf-1 (phospho Ser296) rabbit pAb is a research-use antibody from ELK Biotechnology for studying Raf-1 in WB, IHC, IF, ELISA workflows with listed reactivity for Human, Mouse, Rat. Specifications and supporting documents below should be reviewed when planning the experiment.

Background

This gene is the cellular homolog of viral raf gene (v-raf). The encoded protein is a MAP kinase kinase kinase (MAP3K), which functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Once activated, the cellular RAF1 protein can phosphorylate to activate the dual specificity protein kinases MEK1 and MEK2, which in turn phosphorylate to activate the serine/threonine specific protein kinases, ERK1 and ERK2. Activated ERKs are pleiotropic effectors of cell physiology and play an important role in the control of gene expression involved in the cell division cycle, apoptosis, cell differentiation and cell migration. Mutations in this gene are associated with Noonan syndrome 5 and LEOPARD syndrome 2. [provided by RefSeq, Jul 2008],

Additional antibody information

Specificity Phospho-Raf-1 (S296) Polyclonal Antibody detects endogenous levels of Raf-1 protein only when phosphorylated at S296.
Validation evidence captions Western Blot analysis of 293 cells using Phospho-Raf-1 (S296) Polyclonal Antibody diluted at 1:1000 | Enzyme-Linked Immunosorbent Assay (Phospho-ELISA) for Immunogen Phosphopeptide (Phospho-left) and Non-Phosphopeptide (Phospho-right), using C-RAF (Phospho-Ser296) Antibody | Immunohistochemistry analysis of paraffin-embedded human breast carcinoma, using C-RAF (Phospho-Ser296) Antibody. The picture on the right is blocked with the phospho peptide. | Western blot analysis of lysates from 293 cells treated with PMA 125ng/ml 30' , using C-RAF (Phospho-Ser296) Antibody. The lane on the right is blocked with the phospho peptide.

Key Facts

Product typePrimary Antibody Target: Raf-1
Collaborator ELK Biotechnology
Partner model ES6981
Target Raf-1
Research areas>>EGFR tyrosine kinase inhibitor resistance, >>Endocrine resistance, >>MAPK signaling pathway, >>ErbB signaling pathway, >>Ras signaling pathway, >>Rap1 signaling pathway, >>cGMP-PKG signaling pathway, >>cAMP signaling pathway, >>Chemokine signaling pathway, >>FoxO signaling pathway, >>Sphingolipid signaling pathway, >>Phospholipase D signaling pathway, >>Autophagy - animal, >>mTOR signaling pathway, >>PI3K-Akt signaling pathway, >>Apoptosis, >>Cellular senescence, >>Vascular smooth muscle contraction, >>Axon guidance, >>VEGF signaling pathway, >>Apelin signaling pathway, >>Focal adhesion, >>Gap junction, >>Signaling pathways regulating pluripotency of stem cells, >>Neutrophil extracellular trap formation, >>C-type lectin receptor signaling pathway, >>JAK-STAT signaling pathway, >>Natural killer cell mediated cytotoxicity, >>T cell receptor signaling pathway, >>B cell receptor signaling pathway, >>Fc epsilon RI signaling pathway, >>Fc gamma R-mediated phagocytosis, >>Long-term potentiation, >>Neurotrophin signaling pa
Reactivity Human,Mouse,Rat
Applications WB,IHC,IF,ELISA
ConjugationUnconjugated
Dilution rangeWestern Blot: 1/500 - 1/2000. Immunohistochemistry: 1/100 - 1/300. ELISA: 1/20000. Not yet tested in other applications.
Host Rabbit
Clonality Polyclonal
Isotype IgG
Immunogen The antiserum was produced against synthesized peptide derived from human C-RAF around the phosphorylation site of Ser296. AA range:271-320
Concentration 1 mg/ml
Purification The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
Assay type Antibody validation/application
Expression system In skeletal muscle, isoform 1 is more abundant than isoform 2.
Purity The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
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Application Validation Matrix

3 speciess
ABMIUM Validated™ ABMIUM Verified™ Expected Performance Not suitable
SpeciesWBIHCIFELISAIPFCMChIP
Human
Mouse
Rat
ABMIUM Validated™ WB
Species Dilution Notes
Human Western Blot: 1/500 - 1/2000. Immunohistochemistry: 1/100 - 1/300. ELISA: 1/20000. Not yet tested in other applications.
Mouse
Rat
ABMIUM Validated™ IHC
Species Dilution Notes
ABMIUM Validated™ IF
Species Dilution Notes
ABMIUM Validated™ ELISA
Species Dilution Notes

Specifications

Target
Raf-1
Also known as
RAF1; RAF; RAF proto-oncogene serine/threonine-protein kinase; Proto-oncogene c-RAF; cRaf; Raf-1
Concentration
1 mg/ml
Working concentration
Western Blot: 1/500 - 1/2000. Immunohistochemistry: 1/100 - 1/300. ELISA: 1/20000. Not yet tested in other applications.
Purity
The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
Form / buffer
Liquid
Host species
Rabbit
Clonality
Polyclonal
Isotype
IgG
Conjugation
Unconjugated
Species reactivity
Human,Mouse,Rat
Tested applications
WBIHCIFELISA
Immunogen
The antiserum was produced against synthesized peptide derived from human C-RAF around the phosphorylation site of Ser296. AA range:271-320
Observed band
74kD
Cellular localization
Cytoplasm. Cell membrane. Mitochondrion. Nucleus. Colocalizes with RGS14 and BRAF in both the cytoplasm and membranes. Phosphorylation at Ser-259 impairs its membrane accumulation. Recruited to the cell membrane by the active Ras protein. Phosphorylation at Ser-338 and Ser-339 by PAK1 is required for its mitochondrial localization. Retinoic acid-induced Ser-621 phosphorylated form of RAF1 is predominantly localized at the nucleus.
Dilution range
Western Blot: 1/500 - 1/2000. Immunohistochemistry: 1/100 - 1/300. ELISA: 1/20000. Not yet tested in other applications.
Purification
The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.

Storage & Stability

Long-term storage
'-20°C/1 year
Storage buffer
Liquid in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide.

Compliance & Certifications

ABMIUM
ABMIUM Verified

Collaborator meets ABMIUM quality standards and overall product performance standards have been met.

ISO
ISO 9001

Quality management certification.

RUO
Research Use Only

For research applications. Not for diagnostic use.

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