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B-Raf Proto-Oncogene, Serine/Threonine Kinase Peptide 6

Antibody: CPTC-BRAF-6
Background

Catalog Number:

CPTC-BRAF-6

RRID:

AB_2722029

Target Antigen:

B-Raf Proto-Oncogene, Serine/Threonine Kinase Peptide 6

Isotype:

IgG1

Species:

Mouse Monoclonal Antibody

Last Updated:

12/06/2024

Antigen Recognition(s):

Peptide, Recombinant Full-length, Endogenous

Thematic Panel(s):

Ras Pathway

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Characterization Data [Compare Characterization Data]
  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-BRAF-6 and full length BRAF recombinant protein were measured using surface plasmon resonance. Full-length BRAF recombinant protein was amine coupled onto a Series S CM5 biosensor chip. CPTC-BRAF-6 antibody was used as analyte and was titrated at 64 nM, 16 nM, 4 nM, 1 nM, and 0.25 nM. All binding data were double-referenced and analyzed globally using a 1:1 fitting model. Kinetic constants were difficult to determine. Try to extend the dissociation time. Click image to enlarge

CPTC-BRAF-6 Affinity and Kinetics (Surface Plasmon Resonance)

Result: Positive

Affinity and binding kinetics of CPTC-BRAF-6 and full length BRAF recombinant protein were measured using surface plasmon resonance. Full-length BRAF recombinant protein was amine coupled onto a Series S CM5 biosensor chip. CPTC-BRAF-6 antibody was used as analyte and was titrated at 64 nM, 16 nM, 4 nM, 1 nM, and 0.25 nM. All binding data were double-referenced and analyzed globally using a 1:1 fitting model. Kinetic constants were difficult to determine. Try to extend the dissociation time.


Characterization SOP Files

  IHC Tissue
Click to enlarge image Tissue Microarray core of colon cancer showing immunohistochemical staining for Antibody CPTC-BRAF-6. Click image to enlarge

CPTC-BRAF-6 IHC

Result: Positive

Tissue Microarray core of colon cancer showing immunohistochemical staining for Antibody CPTC-BRAF-6.


Characterization SOP Files

  Immuno-MRM
Click to enlarge image Immuno-MRM chromatogram of CPTC-BRAF-6 antibody with CPTC-BRAF peptide 5 (NCI ID#181) as target Click image to enlarge

CPTC-BRAF-6 iMRM

Result: Positive

Immuno-MRM chromatogram of CPTC-BRAF-6 antibody with CPTC-BRAF peptide 5 (NCI ID#181) as target


Characterization SOP Files

  NCI 60 Protein Array

CPTC-BRAF-6 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


Characterization SOP Files

  Western Blot - Over-expressed transient protein in cell lysate
Click to enlarge image Western Blot using CPTC-BRAF-6 as primary Ab against recombinant BRAF (lane 2). Also included are molecular wt. standards (lane 1). Click image to enlarge

CPTC-BRAF-6 Western Blot

Result: Positive

Western Blot using CPTC-BRAF-6 as primary Ab against recombinant BRAF (lane 2). Also included are molecular wt. standards (lane 1).


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western Blot using CPTC-BRAF-6 as primary Ab against MCF10A-KRas cell lysate (lane 2). Also included are molecular wt. standards (lane 1). Click image to enlarge

CPTC-BRAF-6 Cell Lysate Western Blot

Result: Positive

Western Blot using CPTC-BRAF-6 as primary Ab against MCF10A-KRas cell lysate (lane 2). Also included are molecular wt. standards (lane 1).


Characterization SOP Files

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Antigen: B-Raf Proto-Oncogene, Serine/Threonine Kinase Peptide 6
Background

NCI Identification Number:

00278

Antigen Name:

B-Raf Proto-Oncogene, Serine/Threonine Kinase Peptide 6

CPTC Name:

CPTC-BRAF Peptide 6

Aliases:

B-Raf Proto-Oncogene, Serine/Threonine Kinase; V-Raf Murine Sarcoma Viral Oncogene Homolog B1; V-Raf Murine Sarcoma Viral Oncogene Homolog B; Proto-Oncogene B-Raf; BRAF1; RAFB1; B-Raf Proto-Oncogene Serine/Threonine-Protein Kinase (P94); Murine Sarcoma Viral (V-Raf) Oncogene Homolog B1; Serine/Threonine-Protein Kinase B-Raf; 94 KDa B-Raf Protein; EC 2.7.11.1; B-RAF1; B-Raf; NS7; P94

Function:

This gene encodes a protein belonging to the raf/mil family of serine/threonine protein kinases. This protein plays a role in regulating the MAP kinase/ERKs signaling pathway, which affects cell division, differentiation, and secretion. Mutations in this gene are associated with cardiofaciocutaneous syndrome, a disease characterized by heart defects, mental retardation and a distinctive facial appearance. Mutations in this gene have also been associated with various cancers, including non-Hodgkin lymphoma, colorectal cancer, malignant melanoma, thyroid carcinoma, non-small cell lung carcinoma, and adenocarcinoma of lung. A pseudogene, which is located on chromosome X, has been identified for this gene.

BRAF (B-Raf Proto-Oncogene, Serine/Threonine Kinase) is a Protein Coding gene. Diseases associated with BRAF include Cardiofaciocutaneous Syndrome and Lung Cancer. Among its related pathways are Development Slit-Robo signaling and MAPK Signaling: Oxidative Stress Pathway. GO annotations related to this gene include calcium ion binding and transferase activity, transferring phosphorus-containing groups. An important paralog of this gene is RAF1.

Protein kinase involved in the transduction of mitogenic signals from the cell membrane to the nucleus. May play a role in the postsynaptic responses of hippocampal neuron. Phosphorylates MAP2K1, and thereby contributes to the MAP kinase signal transduction pathway.

Raf kinases, a family of three serine/threonine kinases, are part of the Ras-MAPK signaling cascade and phosphorylate MEK. Upon growth factor stimulation, Raf-1 (c-Raf) is activated by GTP-bound Ras and recruited to the cell membrane. This activation process is tightly regulated.

Chromosomal Localization:

7q34

Accession Number:

NP_004324.2

UniProt Accession Number:

P15056

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

GDGGSTTGLSATPPASLPGSLTNVK

Native Sequence:

Calculated Isoelectric Point:

0

Molecular Weight:

2750

Last Updated:

01/02/2019

Links

Characterization Data

SOPs

No SOPs available.

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