PRODUCT CODE: ET1701-18

A-RAF Recombinant Rabbit Monoclonal Antibody [JJ08-68] (ET1701-18)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

Western blot analysis of A-RAF on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-18, 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.<br />
Positive control: <br />
Lane 1: Hela cell lysate<br />
Lane 2: A431 cell lysate
  • Western blot analysis of A-RAF on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-18, 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.<br />
Positive control: <br />
Lane 1: Hela cell lysate<br />
Lane 2: A431 cell lysate
  • ICC staining of A-RAF in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-18, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 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 A-RAF in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-18, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 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 A-RAF in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-18, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-A-RAF antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-18, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-A-RAF antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-18, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of A-RAF was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-18, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of A-RAF on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-18, 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.
Positive control:
Lane 1: Hela cell lysate
Lane 2: A431 cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

A-RAF Recombinant Rabbit Monoclonal Antibody [JJ08-68] (ET1701-18)

Immunogen

Recombinant protein within human a-raf aa 150-300.

Host

Rabbit

Positive Control

A431, NIH/3T3, Hela, human breast carcinoma tissue, human kidney tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JJ08-68

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

68 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

A-RAF

SYNONYMS

A raf 1 antibody; A Raf proto oncogene serine/threonine protein kinase antibody; ARAF 1 antibody; Araf antibody; ARaf proto oncogene serine/threonine protein kinase antibody; ARAF_HUMAN antibody; ARAF1 antibody; Oncogene Araf1 antibody; Oncogene PKS2 antibody; PKS 2 antibody; PKS antibody; PKS2 antibody; Proto oncogene Pks antibody; Proto-oncogene A-Raf antibody; Proto-oncogene A-Raf-1 antibody; Proto-oncogene Pks antibody; RAFA 1 antibody; RAFA1 antibody; Ras binding protein DA Raf antibody; Serine/threonine-protein kinase A-Raf antibody; v raf murine sarcoma 3611 viral oncogene homolog antibody; v raf murine sarcoma 3611 viral oncogene homolog 1 antibody; v raf oncogene homolog 1 (murine sarcoma 3611 virus) antibody

SEQUENCE SIMILARITIES

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

TISSUE SPECIFICITY

Predominantly in urogenital tissues.

SUBCELLULAR LOCATION

Cytosol.

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. A-Raf, 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 A-Raf is found at highest levels in urogenital tissues and kidney and at lowest level in brain tissue.