c-Raf Antibody [K20J19] | Primary Antibodies

Application: Reactivity: Human, Mouse, Rat, Monkey, Bovine, Pig

Usage Information

Dilution
WB1:1000

Application
WB

Reactivity
Human, Mouse, Rat, Monkey, Bovine, Pig

Source
Mouse Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
75 kDa

Positive Control	A-431 cells; HeLa cells; HT-29 cells; NIH/3T3 cells; NIE-115 cells; H-4-II-E cells; COS-7 cells; Vero cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
c-Raf Antibody [K20J19] detects endogenous levels of total c-Raf protein.

Clone
K20J19

Synonym(s)
RAF proto-oncogene serine/threonine-protein kinase; Proto-oncogene c-RAF (cRaf); Raf-1; RAF1; RAF

Background
c-Raf (Raf-1), together with A-Raf and B-Raf, forms the Raf kinase family of serine/threonine kinases within the TKL group, serving as major effectors recruited by GTP-bound Ras to initiate the MEK-ERK MAPK pathway that orchestrates key cellular processes including proliferation, survival, differentiation, and migration. c-Raf features an N-terminal regulatory region with a Ras-binding domain (RBD) and a cysteine-rich C1 domain for Ras-dependent membrane localization, a hinge region, and a C-terminal kinase domain containing pivotal activation loop phosphorylation sites (Thr491, Ser494, Ser497, Ser499) as well as priming sites Ser338 (phosphorylated by PAK) and Tyr341 (by Src family kinases); its activity is further modulated by inhibitory 14-3-3 binding motifs at Ser259 (Akt-regulated) and Ser621 (AMPK-regulated). Activation of c-Raf is driven by multi-site phosphorylation, which induces conformational changes that facilitate high-affinity MEK1/2 binding and phosphorylation, thereby amplifying ERK1/2 signaling to drive cell cycle progression (via cyclin D), inhibit apoptosis (through Bad phosphorylation), promote cytoskeletal remodeling, and modulate transcription factors such as Elk-1. c-Raf integrates growth factor and Ras inputs into dynamic pathway flux, but sustained MEK-ERK signaling triggers feedback hyperphosphorylation at multiple inhibitory sites (Ser29, Ser43, Ser289, Ser296, Ser301, Ser642), dampening its responsiveness. Dysregulated c-Raf contributes to oncogenesis in Ras-mutant tumors (such as pancreatic and lung cancers), can be paradoxically activated by some inhibitors, and, in contrast to B-Raf^V600E’s constitutive activity in melanoma, requires strict Ras/Src-dependent activation and exhibits lower basal activity.

Background

c-Raf (Raf-1), together with A-Raf and B-Raf, forms the Raf kinase family of serine/threonine kinases within the TKL group, serving as major effectors recruited by GTP-bound Ras to initiate the MEK-ERK MAPK pathway that orchestrates key cellular processes including proliferation, survival, differentiation, and migration. c-Raf features an N-terminal regulatory region with a Ras-binding domain (RBD) and a cysteine-rich C1 domain for Ras-dependent membrane localization, a hinge region, and a C-terminal kinase domain containing pivotal activation loop phosphorylation sites (Thr491, Ser494, Ser497, Ser499) as well as priming sites Ser338 (phosphorylated by PAK) and Tyr341 (by Src family kinases); its activity is further modulated by inhibitory 14-3-3 binding motifs at Ser259 (Akt-regulated) and Ser621 (AMPK-regulated). Activation of c-Raf is driven by multi-site phosphorylation, which induces conformational changes that facilitate high-affinity MEK1/2 binding and phosphorylation, thereby amplifying ERK1/2 signaling to drive cell cycle progression (via cyclin D), inhibit apoptosis (through Bad phosphorylation), promote cytoskeletal remodeling, and modulate transcription factors such as Elk-1. c-Raf integrates growth factor and Ras inputs into dynamic pathway flux, but sustained MEK-ERK signaling triggers feedback hyperphosphorylation at multiple inhibitory sites (Ser29, Ser43, Ser289, Ser296, Ser301, Ser642), dampening its responsiveness. Dysregulated c-Raf contributes to oncogenesis in Ras-mutant tumors (such as pancreatic and lung cancers), can be paradoxically activated by some inhibitors, and, in contrast to B-Raf^V600E’s constitutive activity in melanoma, requires strict Ras/Src-dependent activation and exhibits lower basal activity.

References
https://pubmed.ncbi.nlm.nih.gov/28694330/ https://pubmed.ncbi.nlm.nih.gov/29358316/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%