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B-Raf Proto-Oncogene, Serine/Threonine Kinase Peptide 4

Antibody: CPTC-BRAF-4
Background

Catalog Number:

CPTC-BRAF-4

RRID:

AB_2722027

Target Antigen:

B-Raf Proto-Oncogene, Serine/Threonine Kinase Peptide 4

Isotype:

IgG1

Species:

Mouse Monoclonal Antibody

Last Updated:

12/06/2024

Antigen Recognition(s):

Peptide

Thematic Panel(s):

Ras Pathway

Purchase
Publications
Characterization Data [Compare Characterization Data]
  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-BRAF-4 antibody 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-4 mouse antibody was used as analyte and was titrated at 256 nM, 64 nM, 16 nM, 4 nM, and 1 nM. All binding data were double-referenced and analyzed globally using a 1:1 fitting model. Click image to enlarge

CPTC-BRAF-4 Affinity and Kinetics (Surface Plasmon Resonance)

Result: Positive

Affinity and binding kinetics of CPTC-BRAF-4 antibody 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-4 mouse antibody was used as analyte and was titrated at 256 nM, 64 nM, 16 nM, 4 nM, and 1 nM. All binding data were double-referenced and analyzed globally using a 1:1 fitting model.


Characterization SOP Files

  Immuno-MRM
Click to enlarge image Immuno-MRM chromatogram of CPTC-BRAF-4 antibody with CPTC-BRAF peptide 4 (NCI ID#180) as target Click image to enlarge

CPTC-BRAF-4 iMRM

Result: Positive

Immuno-MRM chromatogram of CPTC-BRAF-4 antibody with CPTC-BRAF peptide 4 (NCI ID#180) as target


Characterization SOP Files

  NCI 60 Protein Array

CPTC-BRAF-4 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 - Tissue or Cell Lysate

CPTC-BRAF-4 Simple Western (cell lysates)

Result: Negative

Simple Western assay using CPTC-BRAF-4 as primary antibody against the whole cell lysates of the cancer cells RPMI-8226, SNB-75, HL-60, NCI/ADR-RES, CCRF-CEM and SR. The target protein BRAF has not been detected in any cell lines. All the cell lines were probed with anti-CytC antibody, and they all show expression of the housekeeping protein at different level.


Characterization SOP Files

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Antibody: CPTC-BRAF-5
Background

Catalog Number:

CPTC-BRAF-5

RRID:

AB_2722028

Target Antigen:

B-Raf Proto-Oncogene, Serine/Threonine Kinase Peptide 4

Isotype:

IgG1

Species:

Mouse Monoclonal Antibody

Last Updated:

12/06/2024

Antigen Recognition(s):

Peptide, Recombinant Full-length, Endogenous

Thematic Panel(s):

Ras Pathway

Purchase
Publications
Characterization Data [Compare Characterization Data]
  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-BRAF-5 and full length BRAF recombinant protein were measured using surface plasmon resonance. CPTC-BRAF-5 mouse antibody was captured on a series S protein G biosensor chip. Full length BRAF recombinant protein was used as analyte and was titrated at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, and 1 nM. All binding data were double-referenced and analyzed globally using a 1:1 fitting model.   Click image to enlarge

CPTC-BRAF-5 Affinity and Kinetics (Surface Plasmon Resonance)

Result: Positive

Affinity and binding kinetics of CPTC-BRAF-5 and full length BRAF recombinant protein were measured using surface plasmon resonance. CPTC-BRAF-5 mouse antibody was captured on a series S protein G biosensor chip. Full length BRAF recombinant protein was used as analyte and was titrated at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, and 1 nM. All binding data were double-referenced and analyzed globally using a 1:1 fitting model.

 


Characterization SOP Files

  IHC Tissue
Click to enlarge image Tissue Microarray core of colon cancer showing immunochistochemical staining for CPTC-BRAF-5. Click image to enlarge

CPTC-BRAF-5 IHC

Result: Positive

Tissue Microarray core of colon cancer showing immunochistochemical staining for CPTC-BRAF-5.


Characterization SOP Files

  Immuno-MRM
Click to enlarge image Immuno-MRM chromatogram of CPTC-BRAF-5 antibody with CPTC-BRAF peptide 4 (NCI ID#180) as target Click image to enlarge

CPTC-BRAF-5 iMRM

Result: Positive

Immuno-MRM chromatogram of CPTC-BRAF-5 antibody with CPTC-BRAF peptide 4 (NCI ID#180) as target


Characterization SOP Files

  NCI 60 Protein Array

CPTC-BRAF-5 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-5 as primary Ab against recombinant BRAF (lane 2). Also included are molecular wt. standards (lane 1). Click image to enlarge

CPTC-BRAF-5 Western Blot

Result: Positive

Western Blot using CPTC-BRAF-5 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-5 as primary Ab against MCF10A-KRas cell lysate (lane 2). Also included are molecular wt. standards (lane 1). Click image to enlarge

CPTC-BRAF-5 Western Blot

Result: Positive

Western Blot using CPTC-BRAF-5 as primary Ab against MCF10A-KRas cell lysate (lane 2). Also included are molecular wt. standards (lane 1).


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Simple Western assay using CPTC-BRAF-5 as primary antibody against the whole cell lysates of the cancer cells RPMI-8226, SNB-75, HL-60, NCI/ADR-RES, CCRF-CEM and SR. The target protein BRAF has been detected in all cell lines, except for HL-60. All the cell lines were probed with anti-CytC antibody, and they all show expression of the housekeeping protein at different level. Click image to enlarge

CPTC-BRAF-5 Simple Western (Cell Lysates)

Result: Positive

Simple Western assay using CPTC-BRAF-5 as primary antibody against the whole cell lysates of the cancer cells RPMI-8226, SNB-75, HL-60, NCI/ADR-RES, CCRF-CEM and SR. The target protein BRAF has been detected in all cell lines, except for HL-60. All the cell lines were probed with anti-CytC antibody, and they all show expression of the housekeeping protein at different level.


Characterization SOP Files

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Antigen: B-Raf Proto-Oncogene, Serine/Threonine Kinase Peptide 4
Background

NCI Identification Number:

00180

Antigen Name:

B-Raf Proto-Oncogene, Serine/Threonine Kinase Peptide 4

CPTC Name:

CPTC-BRAF Peptide 4

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:

RDSSDDWEIPDGQITVGQR

Native Sequence:

Calculated Isoelectric Point:

0

Molecular Weight:

2090

Last Updated:

12/22/2016

Links

Characterization Data

SOPs

No SOPs available.

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