Rabbit Polyclonal Raf-1 antibody (STJ95345)

$99.00$319.00

Reactivity: Human, Mouse, Rat
Applications: WB, IHC, IF
Conjugation: Unconjugated
Supplier: St John’s Laboratory Ltd.

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Rabbit Polyclonal Raf-1 antibody (STJ95345)

Supplier: St John’s Laboratory Ltd.

Recommended applications: WB, IHC, ELISA

Recommended dilution: WB 1:500-1:2000; IHC 1:100-1:300; ELISA 1:5000;

Recommended protocols: check protocols

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Alternative names:

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RAF1 antibody, RAF antibody,|c Raf antibody|C-raf antibody|C-Raf proto-oncogene, serine/threonine kinase antibody|CMD1NN antibody|Craf 1 transforming gene antibody|cRaf antibody|Craf1 transforming gene antibody|EC 2.7.11.1 antibody|kinase Raf1 antibody|Murine sarcoma 3611 oncogene 1 antibody|NS5 antibody|Oncogene MIL antibody|Oncogene RAF1 antibody|OTTHUMP00000160218 antibody|OTTHUMP00000207813 antibody|OTTHUMP00000209389 antibody|Protein kinase raf 1 antibody|Proto-oncogene c-RAF antibody|Raf 1 antibody|Raf 1 proto oncogene serine/threonine kinase antibody|RAF antibody|Raf proto oncogene serine/threonine protein kinase antibody|RAF proto-oncogene serine/threonine-protein kinase antibody|RAF-1 antibody|RAF1 antibody|RAF1_HUMAN antibody|Similar to murine leukemia viral (V-raf-1) oncogene homolog 1 antibody|TRANSFORMING REPLICATION-DEFECTIVE MURINE RETROVIRUS 3611-MSV antibody|v raf 1 murine leukemia viral oncogene homolog 1 antibody|v-raf murine sarcoma viral oncogene homolog 1 antibody|v-raf-1 murine leukemia viral oncogene-like protein 1 antibody|vraf1 murine leukemia viral oncogene homolog 1 antibody|Anti-Raf1 antibody (ab137435)
SCBT cat No: sc-81513|sc-271929|sc-271928|sc-101791|sc-21833|sc-12358|sc-28005|sc-16807|sc-16806|sc-28006|sc-81705|sc-46667|sc-25419|sc-8251|sc-376451|sc-166267|

Name

Raf-1 Polyclonal Antibody

Catalogue No.

STJ95345

Reactivity

Human, Mouse, Rat

Specificity

Raf-1 Polyclonal Antibody detects endogenous levels of Raf-1 protein.

Immunogen

Synthesized peptide derived from Raf-1 at AA range 220-300

Host

Rabbit

Applications

WB, IHC, ELISA

Recommended dilution

WB 1:500-1:2000; IHC 1:100-1:300; ELISA 1:5000;

Clonality

Polyclonal

Conjugation

Unconjugated

Isotype

IgG

Molecular weight

73 kDa

Formulation

Raf-1 Antibody was tube-contained. Liquid in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide.

Concentration

1 mg/ml

Purification

Raf-1 Antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.

Storage

-20 Celsius degree. Avoid repeated freeze/thaw cycles.

Alternative antibody names

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

Database links

Human UniProt/Swiss-Prot:P04049;Mouse UniPort/Swiss-Prot: Q99N57;Rat UniProt/Swiss-Port: P11345;Human Entrez Gene: 5894;Mouse Entrez Gene: 110157;Rat Entrez Gene: Rn.33262

Protein names

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

Protein function

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. / ATP + a protein = ADP + a phosphoprotein. / Zn2+ / Regulation is a highly complex process involving membrane recruitment, protein-protein interactions, dimerization, and phosphorylation/dephosphorylation events. Ras-GTP recruits RAF1 to the membrane, thereby promoting its activation. The inactive conformation of RAF1 is maintained by autoinhibitory interactions occurring between the N-terminal regulatory and the C-terminal catalytic domains and by the binding of a 14-3-3 protein that contacts two phosphorylation sites, Ser-259 and Ser-621. Upon mitogenic stimulation, Ras and PPP2R1A cooperate to release autoinhibition and the subsequent phosphorylation of activating sites: Ser-338, Tyr-341, Thr-491, and Ser-494, yields a fully active kinase. Through a negative feedback mechanism involving MAPK1/ERK2, RAF1 is phosphorylated on Ser-29, Ser-43, Ser-289, Ser-296, Ser-301 and Ser-642 by MAPK1/ERK2, which yields an inactive, desensitized kinase. The signaling-competent conformation of RAF1 is finally re-established by the coordinated action of PIN1, a prolyl isomerase that converts pSer and pThr residues from the cis to the trans conformation, which is preferentially recognized and dephosphorylated by PPP2R1A. Activated by homodimerization and heterodimerization (with BRAF). Also regulated through association with other proteins such as KSR2, CNKSR1/CNK1, PEBP1/RKIP, PHB/prohibitin and SPRY4. PEBP1/RKIP acts by dissociating RAF1 from its substrates MAP2K1/MEK1 and MAP2K2/MEK2. PHB/prohibitin facilitates the displacement of 14-3-3 from RAF1 by activated Ras, thereby promoting cell membrane localization and phosphorylation of RAF1 at the activating Ser-338. SPRY4 inhibits Ras-independent, but not Ras-dependent, activation of RAF1. CNKSR1/CNK1 regulates Src-mediated RAF1 activation.

Protein tissue specificity

In skeletal muscle, isoform 1 is more abundant than isoform 2.

Involvement in disease

Noonan syndrome 5 (NS5) [MIM:611553]: A form of Noonan syndrome, a disease characterized by short stature, facial dysmorphic features such as hypertelorism, a downward eyeslant and low-set posteriorly rotated ears, and a high incidence of congenital heart defects and hypertrophic cardiomyopathy. Other features can include a short neck with webbing or redundancy of skin, deafness, motor delay, variable intellectual deficits, multiple skeletal defects, cryptorchidism, and bleeding diathesis. Individuals with Noonan syndrome are at risk of juvenile myelomonocytic leukemia, a myeloproliferative disorder characterized by excessive production of myelomonocytic cells. . Note: The disease is caused by mutations affecting the gene represented in this entry.; LEOPARD syndrome 2 (LPRD2) [MIM:611554]: A disorder characterized by lentigines, electrocardiographic conduction abnormalities, ocular hypertelorism, pulmonic stenosis, abnormalities of genitalia, retardation of growth, and sensorineural deafness. . Note: The disease is caused by mutations affecting the gene represented in this entry.; Cardiomyopathy, dilated 1NN (CMD1NN) [MIM:615916]: A disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death. . Note: The disease is caused by mutations affecting the gene represented in this entry.

Protein sequence and domain

Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily. / Contains 1 phorbol-ester/DAG-type zinc finger. / Contains 1 protein kinase domain. / Contains 1 RBD (Ras-binding) domain.

Protein post-translational modifications

Phosphorylation at Thr-269, Ser-338, Tyr-341, Thr-491 and Ser-494 results in its activation. Phosphorylation at Ser-29, Ser-43, Ser-289, Ser-296, Ser-301 and Ser-642 by MAPK1/ERK2 results in its inactivation. Phosphorylation at Ser-259 induces the interaction with YWHAZ and inactivates kinase activity. Dephosphorylation of Ser-259 by the complex containing protein phosphatase 1, SHOC2 and M-Ras/MRAS relieves inactivation, leading to stimulate RAF1 activity. Phosphorylation at Ser-338 by PAK1 and PAK7/PAK5 and Ser-339 by PAK1 is required for its mitochondrial localization. Phosphorylation at Ser-621 in response to growth factor treatment stabilizes the protein, possibly by preventing proteasomal degradation. Phosphorylation at Ser-289, Ser-296, Ser-301, Ser-338 and Ser-621 are somehow linked to the methylation potential of cells. Treatment of cells with HGF in the presence of the methylation inhibitor 5′-methylthioadenosine (MTA) results in increased phosphorylation at Ser-338 and Ser-621 and decreased phosphorylation at Ser-296, Ser-301 and Ser-338. Dephosphorylation at Ser-338 by PPP5C results in a activity decrease. / Methylated at Arg-563 in response to EGF treatment. This modification leads to destabilization of the protein, possibly through proteasomal degradation.

Protein cellular localization

Cytoplasm / Cell membrane / Mitochondrion / Nucleus

Research area

All research areas>Kinases and Phosphatases>Raf
(View all antibody categories related to Kinases and Phosphatases)

Note

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Supplier

St John’s Laboratory Ltd.

Product type

Primary antibody

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Immunofluorescence analysis of Rat lung tissue

1: Raf-1 Polyclonal Antibody(red) was diluted at 1:200 (4 degree Celsius,overnight).
2: Cy3 labled Secondary antibody was diluted at 1:300 (room temperature, 50min).
3: Picture B: DAPI(blue) 10min. Picture A:Target. Picture B: DAPI. Picture C: merge of A+B.
 

Immunofluorescence analysis of Rat lung tissue

1: Raf-1 Polyclonal Antibody(red) was diluted at 1:200 (4 degree Celsius,overnight).
2: Cy3 labled Secondary antibody was diluted at 1:300 (room temperature, 50min).
3: Picture B: DAPI(blue) 10min. Picture A:Target. Picture B: DAPI. Picture C: merge of A+B.
 

Immunofluorescence analysis of Rat kidney tissue

1: Raf-1 Polyclonal Antibody(red) was diluted at 1:200 (4 degree Celsius,overnight).
2: Cy3 labled Secondary antibody was diluted at 1:300 (room temperature, 50min).
3: Picture B: DAPI(blue) 10min. Picture A:Target. Picture B: DAPI. Picture C: merge of A+B.
 

Immunofluorescence analysis of Rat kidney tissue

1: Raf-1 Polyclonal Antibody(red) was diluted at 1:200 (4 degree Celsius,overnight).
2: Cy3 labled Secondary antibody was diluted at 1:300 (room temperature, 50min).
3: Picture B: DAPI(blue) 10min. Picture A:Target. Picture B: DAPI. Picture C: merge of A+B.
 

Immunofluorescence analysis of Mouse liver tissue

1: Raf-1 Polyclonal Antibody(red) was diluted at 1:200 (4 degree Celsius,overnight).
2: Cy3 labled Secondary antibody was diluted at 1:300 (room temperature, 50min).
3: Picture B: DAPI(blue) 10min. Picture A:Target. Picture B: DAPI. Picture C: merge of A+B.
 

Immunofluorescence analysis of Mouse liver tissue

1: Raf-1 Polyclonal Antibody(red) was diluted at 1:200 (4 degree Celsius,overnight).
2: Cy3 labled Secondary antibody was diluted at 1:300 (room temperature, 50min).
3: Picture B: DAPI(blue) 10min. Picture A:Target. Picture B: DAPI. Picture C: merge of A+B.
 

Immunohistochemical analysis of paraffin embedded Human liver tissue

1: Raf-1 Polyclonal Antibody was diluted at 1:200 (4 degree Celsius,overnight).
2: Sodium citrate pH 6.0 was used for antibody retrieval (>98 degree Celsius,20min).
3: Secondary antibody was diluted at 1:200 (room temperature, 30min). Negative control was used by secondary antibody only.
 

Immunohistochemical analysis of paraffin embedded Rat liver tissue

1: Raf-1 Polyclonal Antibody was diluted at 1:200 (4 degree Celsius,overnight).
2: Sodium citrate pH 6.0 was used for antibody retrieval (>98 degree Celsius,20min).
3: Secondary antibody was diluted at 1:200 (room temperature, 30min). Negative control was used by secondary antibody only.
 

Immunohistochemical analysis of paraffin embedded Rat kidney tissue

1: Raf-1 Polyclonal Antibody was diluted at 1:200 (4 degree Celsius,overnight).
2: Sodium citrate pH 6.0 was used for antibody retrieval (>98 degree Celsius,20min).
3: Secondary antibody was diluted at 1:200 (room temperature, 30min). Negative control was used by secondary antibody only.


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