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Phospho-Raf1-S621 Rabbit pAb (AP0087)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human

Western blot analysis of lysates from HeLa and 293T cells, using Phospho-Raf1-S621 Rabbit pAb (AP0087) at 1:1000 dilution. HeLa cells were treated by EGF (100ng/mL) for 30 minutes after serum-starvation overnight. 293T cells were treated by PMA/TPA (200nM) for 30 minutes or treated by EGF (25μg/mL) for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% BSA.

ABclonal:Immunoprecipitation - Phospho-Raf1-S621 Rabbit pAb (AP0087)

Immunoprecipitation analysis of 200 μg extracts of HeLa cells, using 3 μg Phospho-Raf1-S621 pAb (AP0087). Western blot was performed from the immunoprecipitate using Phospho-Raf1-S621 pAb (AP0087) at a dilution of 1:1000. HeLa cells were treated by EGF (100 ng/mL) at 37℃ for 30 minutes after serum-starvation overnight.

Overview

Product name	Phospho-Raf1-S621 Rabbit pAb
Catalog No.	AP0087
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG

Background

This gene is the cellular homolog of viral raf gene (v-raf). The encoded protein is a MAP kinase kinase kinase (MAP3K), which functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Once activated, the cellular RAF1 protein can phosphorylate to activate the dual specificity protein kinases MEK1 and MEK2, which in turn phosphorylate to activate the serine/threonine specific protein kinases, ERK1 and ERK2. Activated ERKs are pleiotropic effectors of cell physiology and play an important role in the control of gene expression involved in the cell division cycle, apoptosis, cell differentiation and cell migration. Mutations in this gene are associated with Noonan syndrome 5 and LEOPARD syndrome 2.

Immunogen information

Immunogen	A synthetic phosphorylated peptide around S621 of human Phospho-Raf1-S621 (NP_002871.1).
Sequence	SASEP
Gene ID
Swiss Prot
Synonyms	NS5; CRAF; Raf-1; c-Raf; CMD1NN; Phospho-Raf1-S621
Calculated MW	73kDa
Observed MW	73kDa

Applications

Reactivity	Human
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:500 - 1:2000 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.02% sodium azide, 50% glycerol, pH7.3.
Key application	Western blotting Immunoprecipitation
Positive samples	HeLa treated by EGF, 293T
Cellular location	Cell membrane, Cytoplasm, Mitochondrion, Nucleus