A-RAF Recombinant Rabbit Monoclonal Antibody [JJ08-68]
Catalog# ET1701-18
A-RAF Recombinant Rabbit Monoclonal Antibody [JJ08-68]
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WB
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IF-Tissue
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IHC-P
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FC
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Human
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Mouse
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unconjugated
Overview
Product Name
A-RAF Recombinant Rabbit Monoclonal Antibody [JJ08-68]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human A-RAF aa 146-310 / 606.
Species Reactivity
Human, Mouse
Validated Applications
WB, IF-Tissue, IHC-P, FC
Molecular Weight
Predicted band size: 68 kDa
Positive Control
Hela cell lysate, A431 cell lysate, human kidney tissue, human breast carcinoma tissue, HeLa.
Conjugation
unconjugated
Clone Number
JJ08-68
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:2,000
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IF-Tissue
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1:50-1:200
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IHC-P
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1:50-1:200
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FC
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1:100
Target
Function
Serine/threonine-protein kinase A-Raf or simply A-Raf is an enzyme that in humans is encoded by the ARAF gene. A-Raf is a member of the Raf kinase family of serine/threonine-specific protein kinases. Compared to the other members of this family (Raf-1 and B-Raf), very little is known about A-Raf. It seems to share many of the properties of the other isoforms, but its biological functions are not as thoroughly researched. All three Raf proteins are involved in the MAPK signaling pathway. There are several ways A-Raf is different from the other Raf kinases. A-Raf is the only steroid hormone-regulated Raf isoform. In addition, the A-Raf protein has amino acid substitutions in a negatively charged region upstream of the kinase domain (N-region). This could be responsible for its low basal activity.
Background References
1. K hler, M. et al. 2016. Activation loop phosphorylation regulates B-Raf in vivo and transformation by B-Raf mutants. The EMBO journal. 35: 143-61.
2. Jiao, Q. et al. 2014. Xanthoceraside induces apoptosis in melanoma cells through the activation of caspases and the suppression of the IGF-1R/Raf/MEK/ERK signaling pathway. Journal of medicinal food. 17: 1070-8.
Sequence Similarity
Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily.
Tissue Specificity
Predominantly in urogenital tissues.
Subcellular Location
Cytosol.
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
ExpandImages
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☑ Knockdown (KD)
Western blot analysis of A-RAF on different lysates with Rabbit anti-A-RAF antibody (ET1701-18) at 1/1,000 dilution.
Lane 1: A549-WT cell lysate
Lane 2: A549-KD A-RAF cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 68 kDa
Observed band size: 68 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-18) at 1/1,000 dilution was used in 2% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
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 -
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.
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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.
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Application: Flow Cytometry (Intra)
Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: ET1701-18(1/100, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"