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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 5777, (A:1 μg/mL; B:2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-chicken 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 Western Blot Validation in Human Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: B-raf 5777, (0.5 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Figure 1 Western Blot Validation in Human Brain Tissue
Loading: 15 μg of lysates per lane.Antibodies: B-raf 5777, (A:1 μg/mL; B:2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-chicken IgG HRP conjugate at 1:10000 dilution.
IAV
Trial Size

B-raf Antibody

CATALOG NUMBER: 5777

Clonality:
Polyclonal
Tested Applications:
ELISA, WB
Host Species:
Chicken
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Chicken
Species Reactivity:
Human, Mouse, Rat
Homology:
Predicted species reactivity based on immunogen sequence: Chicken: (93%)
Immunogen:
Anti-B-raf antibody (5777) was raised against a peptide corresponding to 18 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, WB
Application Note:
WB: 0.5-2 μg/mL.
Antibody validated: Western Blot in human and mouse samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1303 - Human Brain Tissue Lysate
Positive Control 2:
Cat. No. 1210 – 293 Cell Lysate
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 has one isoform (712aa, 80kD). 5777 can detect in human B-raf and mouse isoform 1.
Purification:
B-raf Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgY
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. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
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:

5777

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
Shipping Info

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