Key Product Details

Species Reactivity

Human

Applications

Western Blot, Simple Western

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all B-Raf Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human B-Raf
Met1-His766
Accession # P15056

Specificity

Detects human B-Raf in Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human B‑Raf Antibody

Detection of Human B-Raf antibody by Western Blot.

Detection of Human B-Raf by Western Blot.

Western blot shows lysates of T47D human breast cancer cell line, MDA-MB-453 human breast cancer cell line, Daudi human Burkitt's lymphoma cell line, MOLT-4 human acute lymphoblastic leukemia cell line and U266 human myeloma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human B-Raf Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3424) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for B-Raf at approximately 95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human B‑Raf antibody by Simple WesternTM.

Detection of Human B‑Raf by Simple WesternTM.

Simple Western lane view shows lysates of T47D human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for B-Raf at approximately 95 kDa (as indicated) using 10 µg/mL of Goat Anti-Human B-Raf Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3424) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human B‑Raf antibody by Simple WesternTM.

Detection of Human B‑Raf by Simple WesternTM.

Simple Western lane view shows lysates of MOLT‑4 human acute lymphoblastic leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for B‑Raf at approximately 95 kDa (as indicated) using 10 µg/mL of Goat Anti-Human B‑Raf Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3424) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human B-Raf by Western Blot

Detection of Human B-Raf by Western Blot

Changes in ERK1/2 and MEK in response to PLX4032. (A) Melanoma cell strains were treated with DMSO for 4 h (0), or with PLX4032 (1 μM) for 4 and 8 h. The panels show Western blots probed with antibodies to phosph-ERK1/2 Thr202/Tyr204 mAb (pERK), ERK1/2 (ERK), phospho-MEK1/2 (pMEK), MEK1/2 (MEK), and actin as a loading control. The mutation status of BRAF, NRAS, PTEN and beta -catenin ( beta CAT) are indicated at the top. (B) Western blot analyses of ERK1/2 inactivation/activation after short-term incubation with PLX4032 (1 μM). (C) pERK1/2 and ERK1/2 in supernatant (Sup) and particulate (Part) fractions. (D) Changes in pERK1/2 activation after treatments with increasing concentration of PLX4032, or DMSO for 1 h. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20149136), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human B‑Raf Antibody

Application
Recommended Usage

Simple Western

10 µg/mL
Sample: T47D human breast cancer cell line and MOLT‑4 human acute lymphoblastic leukemia cell line

Western Blot

1 µg/mL
Sample: T47D human breast cancer cell line, MDA-MB-453 human breast cancer cell line, Daudi human Burkitt's lymphoma cell line, MOLT-4 human acute lymphoblastic leukemia cell line and U266 human myeloma cell line

Reviewed Applications

Read 1 review rated 4 using AF3424 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: B-Raf

The Raf serine/threonine kinases are effectors of Ras that function as MAP3Ks in the ERK phosphorylation cascade. Mammals express three Raf proteins: Raf-1 (C‑Raf); A-Raf; and B-Raf, found at high levels in cerebrum and testes. Mice with a targeted disruption of the B-Raf gene die of vascular defects during mid-gestation. B-Raf mutations have been found in two-thirds of malignant melanomas, with the single substitution V599E in the kinase domain the most frequent occurrence. B-Raf activation in melanomas results in BIM phosphorylation and inhibition of apoptosis.

Long Name

v-Raf Murine Sarcoma Viral Oncogene Homolog B1

Alternate Names

BRaf, BRAF1, RAFB1

Entrez Gene IDs

673 (Human); 114486 (Rat)

Gene Symbol

BRAF

UniProt

P15056

Additional B-Raf Products

Product Documents for Human B‑Raf Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human B‑Raf Antibody

For research use only

Related Research Areas

Cancer Biomarkers
Intracellular Kinases

Citations for Human B‑Raf Antibody

Customer Reviews for Human B‑Raf Antibody (1)

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  • Human B-Raf Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Cancer cell lysates
    Species: Human
    Verified Customer | Posted 02/07/2018

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