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<strong>Figure 1 Western Blot Validation in Human Brain Tissue</strong><br>Loading: 15 μg of lysates per lane.Antibodies: B-raf 7379, (A:1 μg/mL; B:2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: B-raf 5777, (0.5 μg/mL), B-raf 7379, (0.5 μg/mL) and GAPDH (0.02 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-chicken (for 5777) and goat anti-rabbit (for 7379) IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Immunohistochemistry Validation of B-raf in Human Small Intestine Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded Human Small Intestine Tissue using anti-B-raf antibody (7379) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 4 Immunofluorescence Validation of B-raf in Human Small Intestine Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Small Intestine Tissue labeling B-raf with 7379 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Figure 1 Western Blot Validation in Human Brain Tissue
Loading: 15 μg of lysates per lane.Antibodies: B-raf 7379, (A:1 μg/mL; B:2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
IAV
Trial Size

B-raf Antibody

CATALOG NUMBER: 7379

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Homology:
Predicted species reactivity based on immunogen sequence: Rat: (100%), Chicken: (100%)
Immunogen:
Anti-B-raf antibody (7379) was raised against a peptide corresponding to 19 amino acids near the center of human B-raf.
The immunogen is located within amino acids 330 - 380 of B-raf.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
Application Note:
WB: 0.5-2 μg/mL; IHC: 2.5 μg/mL; IF: 20 μg/mL.
Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry in human samples and Immunofluorescence in human samples. All other applications and species not yet tested.
Specificity:
This antibody will not cross-react with C-raf.
Positive Control 1:
Cat. No. 1303 - Human Brain Tissue Lysate
Positive Control 2:
Cat. No. 11-801 - Human Small Intestine Tissue Slide
Positive Control 3:
Cat. No. 1224 – Daudi Cell Lysate
Positive Control 4:
Cat. No. 1204 – K562 Cell Lysate
Predicted Molecular Weight:
Predicted: 94kD
Observed: 94 kD
Validation:

Independent Antibody Validation in Cell lines (Figure 2) shows similar B-raf expression profile in human and mouse cell lines detected by two independent anti-B-raf antibodies that recognize different epitopes, 5777 against central domain and 7379 against a slight different central domain.  B-raf proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies. 

Isoforms:
Human B-raf has one isoform (766aa, 84kD). Mouse B-raf has 2 isoforms, including isoform 1 (804aa, 89kD) and isoform 2 (329aa, 37kD). Rat B-raf a has one isoform (712aa, 80kD). 7379 can detect in human B-raf and mouse isoform 1.
Purification:
B-raf Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
B-raf Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
B-raf antibody can be stored at 4°C for three months and -20°C, stable for up to one year.
Ncbi Official Symbol:
BRAF
Additional Names:
B-raf Antibody: NS7, BRAF1, RAFB1, B-RAF1, Proto-oncogene B-Raf
Protein Accession Number:
P15056
Protein Gi Number:
50403720
Ncbi Gene Id Number:
673
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
B-raf Antibody: B-raf belongs to the raf/mil family of serine/threonine protein kinases and plays a role in regulating the MAP kinase/ERKs signaling pathway, which affects cell division, differentiation, and secretion. The Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt pathways interact with each other to regulate growth and in some cases tumorigenesis. Mutations in B-raf have been associated with several cancers, including non-Hodgkin lymphoma, colorectal cancer, malignant melanoma, thyroid carcinoma, non-small cell lung carcinoma, and adenocarcinoma of lung, leading to speculation on the possibility of B-raf as a therapeutic target for treating cancers. Mutations in this gene have also been associated with cardiofaciocutaneous syndrome (CFCS), a disease characterized by heart defects, mental retardation and a distinctive facial appearance.
Background Reference 1:
McCubrey et al. Biophys. Acta 2007; 1773:1263-84.
Background Reference 2:
Madhunapantula and Robertsonet al. Cancer Res.2008; 68:5-8.
Background Reference 3:
Sarkozy et al. Hum. Mutat.2009; 30:695-702.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

7379

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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