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PRODUCT CODE: ET1701-20

Phospho-B Raf (T401) Recombinant Rabbit Monoclonal Antibody [JJ08-72] (ET1701-20)

  • Recombinant

Applications

  • WB

  • ICC

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of Phospho-B Raf(T401) on mouse testis tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-20, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of Phospho-B Raf(T401) on mouse testis tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-20, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of Phospho-B Raf(T401) on PC-12 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-20, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
  • ICC staining of Phospho-B Raf(T401) in PC-3M cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-20, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-B Raf(T401) in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-20, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-B Raf(T401) in PC-12 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-20, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Western blot analysis of Phospho-B Raf(T401) on mouse testis tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-20, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Phospho-B Raf (T401) Recombinant Rabbit Monoclonal Antibody [JJ08-72] (ET1701-20)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding thr401 of human b raf aa 371-420 / 766.

Host

Rabbit

Modification

Phospho

Modification Site

T401

Positive Control

PC-3M, MCF-7, PC-12, mouse testis tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JJ08-72

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

84 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:5,000

  • ICC

  • 1:100-1:500

TARGET

UNIPROT #

P15056

PROTEIN NAME

Phospho-B Raf (T401)

SYNONYMS

FLJ95109 antibody; 94 kDa B raf protein antibody; B raf 1 antibody; B Raf proto oncogene serine threonine protein kinase antibody; B Raf proto oncogene, serine/threonine kinase antibody; B RAF1 antibody; B-Raf proto-oncogene serine/threonine-protein kinase (p94) antibody; BRAF 1 antibody; BRAF antibody; BRAF_HUMAN antibody; BRAF1 antibody; cRmil antibody; MGC126806 antibody; MGC138284 antibody; Murine sarcoma viral (v-raf) oncogene homolog B1 antibody; Murine sarcoma viral v raf oncogene homolog B1 antibody; NS7 antibody; Oncogen BRAF antibody; oncogene BRAF1 antibody; p94 antibody; Proto-oncogene B-Raf antibody; Proto-oncogene c-Rmil antibody; RAFB 1 antibody; RAFB1 antibody; RMIL antibody; Serine/threonine-protein kinase B-raf antibody; v raf murine sarcoma viral oncogene homolog B antibody; v raf murine sarcoma viral oncogene homolog B1 antibody; v-Raf murine sarcoma viral oncogene homolog B1 antibody

SEQUENCE SIMILARITIES

Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily.

TISSUE SPECIFICITY

Brain and testis.

POST-TRANSLATIONAL MODIFICATION

Phosphorylation at Ser-365 by SGK1 inhibits its activity.; Methylation at Arg-671 decreases stability and kinase activity.; Ubiquitinated by RNF149; which leads to proteasomal degradation. Polyubiquitinated at Lys-578 in response to EGF.

SUBCELLULAR LOCATION

Nucleus, Cytoplasm, Cell membrane.

FUNCTION

Several serine/threonine protein kinases have been implicated as intermediates in signal transduction pathways. These include ERK/MAP kinases, ribosomal S6 kinase (Rsk) and Raf-1. Raf-1 is a cytoplasmic protein with intrinsic serine/threonine activity. It is broadly expressed in nearly all cell lines tested to date and is the cellular homolog of v-Raf, the product of the transforming gene of the 3611 strain of murine sarcoma virus. The unregulated kinase activity of the v-Raf protein has been associated with transformation and mitogenesis, while the activity of Raf-1 is normally suppressed by a regulatory N-terminal domain. Raf-A, a second member of the Raf gene family of serine/threonine protein kinases, exhibits substantial homology to Raf-1 within the kinase domain of the two molecules, but less homology elsewhere. Expression of Raf-B is highly restricted, with highest levels in the cerebrum and testis.