c-RAF (Phospho-S621) Rabbit Monoclonal Antibody(C1513)

MSDS

c-RAF (Phospho-S621) Rabbit Monoclonal Antibody(C1513)

Key features and details

Recombinant rabbit monoclonal antibody to c-RAF (Phospho-S621)

Target: c-RAF (Phospho-S621)
Source/Host: Rabbit
Reactivity: Human
Clonality: Monoclonal
Applications: WB, IF/ICC
Conjugation: Unconjugated
Storage: at-20°C

Brand:

CAT.NO. : AMA01125

US$ Please choose

Size:

50μL 100μL

Trail, Bulk size or Custom requests Please contact us

Product Details

Background

Serine/threonine-protein kinase that acts as a regulatory link between the membrane-associated Ras GTPases and the MAPK/ERK cascade, and this critical regulatory link functions as a switch determining cell fate decisions including proliferation, differentiation, apoptosis, survival and oncogenic transformation. RAF1 activation initiates a mitogen-activated protein kinase (MAPK) cascade that comprises a sequential phosphorylation of the dual-specific MAPK kinases (MAP2K1/MEK1 and MAP2K2/MEK2) and the extracellular signal-regulated kinases (MAPK3/ERK1 and MAPK1/ERK2). The phosphorylated form of RAF1 (on residues Ser-338 and Ser-339, by PAK1) phosphorylates BAD/Bcl2-antagonist of cell death at 'Ser-75'. Phosphorylates adenylyl cyclases: ADCY2, ADCY5 and ADCY6, resulting in their activation.

Application

To ensure optimal assay performance, AREX recommends conducting reagent titration tailored to each testing system for optimal detection results.

WB	1:500 - 1:1000
IF/ICC	1:50 - 1:200

*Results are sample-specific. Please refer to your local assay conditions and test parameters for reference.

Overview

Description	Recombinant rabbit monoclonal antibody to c-RAF (Phospho-S621)
Specificity	Recognizes endogenous levels of c-RAF protein only when phosphorylated at S621
Antibody Type	Primary antibody, Recombinant
Imnunogen	KLH-conjugated synthetic phosphopeptide corresponding to residues surrounding S621 of human c-RAF protein. The exact sequence is proprietary.
Purification	The antibody was purified by immunogen affinity chromatography.
Molecular Weight	Predicted: 73 kD; Observed: 73 kD
Form/Buffer	Liquid in PBS, pH 7.4, containing 50% glycerol, 0.2% BSA and 0.01% sodium azide.
Alternative Names	RAF; RAF proto-oncogene serine/threonine-protein kinase; Proto-oncogene c-RAF; cRaf; Raf-1
Gene Symbol	RAF1
Entrez Gene	5894(Human)
SwissProt	P04049(Human)

*AREX continuously optimizes its products. Webpage content may not reflect the latest updates. For inquiries, please contact info@arexbio.com or your local distributor. Clone Number, Reactivity, Source/Host and Clonality can be found in the product name and Key Features section above.

Data

Western blot analysis of c-RAF (Phospho-S621) expression in PC3 (A), MCF7 (B) whole cell lysates. (Predicted band size: 73 kD; Observed band size: 73 kD)

Immunofluorescent analysis of c-RAF (Phospho-S621) staining in MCF7 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 °C in a hidified chamber. Cells were washed with PBST and incubated with a AREX® Fluor 488 -conjugated secondary antibody (green) in PBS at room temperature in the dark. Phalloidin - AREX® Fluor 594 was used to stain Actin filaments (red). DAPI was used to stain the cell nuclei (blue).

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Note

For Research Use Only. Not for use in diagnostic procedures.

FAQs

What are the main types of research antibodies and how do they differ?

Research antibodies are mainly divided into monoclonal antibodies and polyclonal antibodies. Monoclonal antibodies typically offer higher specificity and better batch-to-batch consistency, while polyclonal antibodies often provide stronger affinity but may show more variation between batches. The choice depends on your specific experimental needs.

How can I tell if a research antibody is suitable for my experiment?

It is recommended to carefully review the product datasheet for validated applications, species reactivity, recommended dilutions, and published references. For new antibodies, performing a small-scale validation with positive control samples is usually helpful.

Can improper storage of research antibodies affect experimental results?

Yes. Antibodies are sensitive to temperature, repeated freeze-thaw cycles, and contamination. Improper storage may lead to reduced activity, increased background, or weaker signals. It is best to follow the storage instructions provided in the product datasheet.

Why doesn’t the recommended dilution in the datasheet work well in my experiment?

The recommended dilution is based on the supplier’s test conditions. Factors such as sample type, fixation method, and detection system in your lab can influence the optimal working concentration. Performing a dilution series optimization in your own system is often necessary.

What precautions should I take when using a newly purchased research antibody for the first time?

It is advisable to briefly centrifuge the antibody (especially concentrated or lyophilized ones), then perform a small-scale pilot experiment using the recommended conditions. Recording the batch number and usage date is also helpful for future tracking.

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