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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:

07/02/2025

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: High Binding

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 Tissue

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 Simple Western Blot (Overexpressed Lysate)

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 - Over-expressed transient protein in cell lysate

CPTC-BRAF-6 Western Blot- Over-expressed Lysate

Result: Negative

Western Blot of CPTC-BRAF-6 against the over-expressed lysate of BRAF. Expected MW is 84 KDa. The antibody did not detect the target protein.


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 Simple Western Blot (Cell Lysate)

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

  Western Blot - Tissue or Cell Lysate

CPTC-BRAF-6 Western Blot- Cell Lysates

Result: Negative

Western blot using CPTC-BRAF-6 as primary antibody against the whole lysates of RPMI-8226, SNB-75, HL-60, NCI/ADR-RES, TK-10, and KM12. The antibody did not detect the target protein (~84 KDa) in any of the tested cell lysates.


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; BRAF1; BRAF-1; V-Raf Murine Sarcoma Viral Oncogene Homolog B1; V-Raf Murine Sarcoma Viral Oncogene Homolog B; Serine/Threonine-Protein Kinase B-Raf; B-Raf Serine/Threonine-Protein; Proto-Oncogene B-Raf; RAFB1; B-Raf Proto-Oncogene Serine/Threonine-Protein Kinase (P94); V-Raf Murine Sarcoma Viral Oncogene-Like Protein B1; Murine Sarcoma Viral (V-Raf) Oncogene Homolog B1; Serine/Threonine-Protein Kinase B-Raf Variant; Non-Specific Serine/Threonine Protein Kinase; Uncharacterized Protein BRAF; 94 KDa B-Raf Protein; BRAF Protein; B-RAF1; B-Raf; NS7; P94

Function:

This gene encodes a protein belonging to the RAF family of serine/threonine protein kinases. This protein plays a role in regulating the MAP kinase/ERK signaling pathway, which affects cell division, differentiation, and secretion. Mutations in this gene, most commonly the V600E mutation, are the most frequently identified cancer-causing mutations in melanoma, and have been identified in various other cancers as well, including non-Hodgkin lymphoma, colorectal cancer, thyroid carcinoma, non-small cell lung carcinoma, hairy cell leukemia and adenocarcinoma of lung. Mutations in this gene are also associated with cardiofaciocutaneous, Noonan, and Costello syndromes, which exhibit overlapping phenotypes. A pseudogene of this gene has been identified on the X chromosome. BRAF mutations are found to be recurrent in many cancer types. Of these, the mutation of valine 600 to glutamic acid (V600E) is the most prevalent. V600E has been determined to be an activating mutation, and cells that harbor it, along with other V600 mutations are sensitive to the BRAF inhibitor dabrafenib. It is also common to use MEK inhibition as a substitute for BRAF inhibitors, and the MEK inhibitor trametinib has seen some success in BRAF mutant melanomas. BRAF mutations have also been correlated with poor prognosis in many cancer types, although there is at least one study that questions this conclusion in papillary thyroid cancer. Oncogenic BRAF mutations are divided into three categories that determine their sensitivity to inhibitors. Class 1 BRAF mutations (V600) are RAS-independent, signal as monomers and are sensitive to current RAF monomer inhibitors. Class 2 BRAF mutations (K601E, K601N, K601T, L597Q, L597V, G469A, G469V, G469R, G464V, G464E, and fusions) are RAS-independent, signaling as constitutive dimers and are resistant to vemurafenib. Such mutants may be sensitive to novel RAF dimer inhibitors or MEK inhibitors. Class 3 BRAF mutations (D287H, V459L, G466V, G466E, G466A, S467L, G469E, N581S, N581I, D594N, D594G, D594A, D594H, F595L, G596D, and G596R) with low or absent kinase activity are RAS-dependent and they activate ERK by increasing their binding to activated RAS and wild-type CRAF. Class 3 BRAF mutations coexist with mutations in RAS or NF1 in melanoma may be treated with MEK inhibitors. In epithelial tumors such as CRC or NSCLC may be effectively treated with combinations that include inhibitors of receptor tyrosine kinase.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.BRAF (B-Raf Proto-Oncogene, Serine/Threonine Kinase) is a Protein Coding gene. Diseases associated with BRAF include Cardiofaciocutaneous Syndrome 1 and Noonan Syndrome 7. Among its related pathways are In Progress: Dabrafenib Pathway, Pharmacokinetics/Pharmacodynamics and Signalling to p38 via RIT and RIN. An important paralog of this gene is RAF1.

Chromosomal Localization:

7q34

Accession Number:

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:

07/01/2026

Links

Characterization Data

SOPs

No SOPs available.

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