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Purified anti-B-Raf Antibody

Pricing & Availability
Clone
W24048D (See other available formats)
Regulatory Status
RUO
Other Names
BRAF, V-Raf Murine Sarcoma ViralOncogene Homolog B1, Serine/threonine-protein k
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
1. W24048D_PURE_B-Raf_1_100225
Whole cell extracts (15 µg total protein per lane) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-B-Raf (clone W24048D) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • 1. W24048D_PURE_B-Raf_1_100225
    Whole cell extracts (15 µg total protein per lane) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-B-Raf (clone W24048D) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • 2. W24048D_PURE_B-Raf_2_100225
    Whole cell extracts (15 µg total protein per lane) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-B-Raf (clone W24048D) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • 3. W24048D_PURE_B-Raf_3_100225
    IHC staining with Purified anti-B-Raf (clone W24048D) on formalin-fixed paraffin-embedded human testis tissue. Following antigen retrieval using 1X Citrate Buffer (Cat. No. 420902), the tissue was incubated with Purified anti-B-Raf (clone W24048D) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 10X (panels A and B) and 40X (panels C and D) objective and merged (panels B and D). Scale bar: 50 µm
  • 4. W24048D_PURE_B-Raf_4_100225
    IHC staining with Purified anti-B-Raf (clone W24048D) on formalin-fixed paraffin-embedded human prostate cancer tissue. Following antigen retrieval using 1X Citrate Buffer (Cat. No. 420902), the tissue was incubated with (panel A) or without (panel C) Purified anti-B-Raf (clone W24048D) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 40X objective and merged (panels B and D). Scale bar: 50 µm
  • 5. W24048D_PURE_B-Raf_5_100225
    HAP1 WT cells (positive cell line, filled histogram) and HAP1 B-Raf KO cells (negative cell line, open histogram) were fixed and permeabilized using Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803) and intracellularly stained with Purified anti-B-Raf (clone W24048D) or Purified Rat IgG2a, κ Isotype Control (open histogram, dashed line, representative of both the positive and negative cell lines) (Cat No. 400502) followed by Alexa Fluor® 647 anti-rat IgG (Cat. No. 405416).
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648886 25 µg 124€
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648887 100 µg 324€
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Description

B-Raf is one of three proteins in the RAF family of serine/threonine protein kinases. B-Raf regulates the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway. Like the other two RAF kinases, A-Raf and C-Raf, B-Raf relays extracellular signals to the nucleus through the MAPK/ERK pathway and regulates cell division, differentiation, and secretion. RAF proteins share similar protein domains and motifs, including multiple protein binding domains such as Ras, lipid, and the 14-3-3 binding motif. RAF proteins mainly reside in the cytoplasm in an inactive state by binding to 14-3-3 proteins. RAFs translocate to the membrane compartment upon activation and binding with RAS proteins. Similar to other RAF kinases, B-Raf phosphorylates MAP2K1 (MEK) proteins to activate the MAPK/ERK pathway. Mutations in the gene that encodes B-Raf are highly correlated with inherited cardiofaciocutaneous syndrome and various types of cancers. The B-Raf V600E mutation results in a constitutively active kinase leading to aberrant cell proliferation and invasion. Such mutations are linked to hairy cell leukemia, melanoma, colon cancer, and non-small-cell lung cancer. B-Raf inhibitors are considered as anticancer targets for metastatic melanoma, kidney cancer, and liver cancer.

Product Details
Quality Statement

This product was carefully developed for performance and specificity using the following products from Horizon Discovery, a Revvity company:
HAP1 B-Raf knockout cell lines (Horizon Cat. No. HZGHC000002c001)
HAP1 parental cell lines (Horizon Cat. No. C631)

Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Recombinant fragment of human B-Raf
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
IHC-P, ICFC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.25 - 1.0 µg/mL. For immunohistochemistry, a concentration range of 1.0 - 10.0 µg/mL is suggested. For flow cytometric staining, the suggested use of this reagent is ≤ 0.125 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Additional Product Notes

For use in immunohistochemistry on formalin-fixed paraffin-embedded tissue (IHC-P), it is recommended to perform antigen retrieval using either Citrate Buffer, 10X (Cat. No. 420902) or Tris-EDTA pH 9.0 Antigen Retrieval Buffer (10X) (Cat. No. 422704).

For use in intracellular flow cytometry (ICFC), it is recommended to fix/permeabilize with either Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803), True-Phos™ Perm Buffer (Cat. No. 425401) or True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401).

RRID
AB_3720627 (BioLegend Cat. No. 648886)
AB_3720627 (BioLegend Cat. No. 648887)

Antigen Details

Structure
B-Raf is a 766 amino acid protein with a predicted molecular weight of 84.4 kD. It forms homodimers and heterodimers with C-Raf.
Distribution

Cytoplasm, membrane, and nucleus

Function
Phosphorylates MAP2K1 and activates the MAPK/ERK pathway
Interaction
Regulates the MAPK and ERK pathways; interacts with PFKFB2
Biology Area
Cancer Biomarkers, Cardiovascular Biology, Cell Cycle/DNA Replication, Cell Proliferation and Viability, Neuroscience, Synaptic Biology
Molecular Family
Postsynaptic proteins, Phospho-Proteins, Protein Kinases/Phosphatase
Antigen References
  1. Davies H, et al. 2002. Nature. 17(6892):949-54.
  2. Wajapeyee N, et al. 2008. Cell. 132(3):363-74.
  3. Tiacci E, et al. 2011. N. Engl. J. Med. 364(24):2305-15.
  4. Dankner M, et al. 2018. Oncogene. 37(24):3183-3199.
Gene ID
673 View all products for this Gene ID
UniProt
View information about B-Raf on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 10/02/2025

For Research Use Only. Not for diagnostic or therapeutic use.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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