RAF1 Antikörper (V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1) (AA 25-60)

Details for Product anti-RAF1 Antibody No. ABIN733208, Anbieter: Anmelden zum Anzeigen
Antigen
  • c-raf
  • craf
  • raf1
  • RAF1
  • raf
  • CRAF
  • NS5
  • Raf-1
  • c-Raf
  • 6430402F14Rik
  • AA990557
  • BB129353
  • Craf1
  • D830050J10Rik
  • v-Raf
  • AFR1
  • RAF
  • Afr-1
  • Afr1
  • Raf
  • mKIAA0854
  • v-raf-1 murine leukemia viral oncogene homolog 1a
  • v-raf-1 murine leukemia viral oncogene homolog 1
  • raf protein
  • v-raf-leukemia viral oncogene 1
  • zinc fingers and homeoboxes 2
  • raf1a
  • RAF1
  • raf1
  • LOC100341921
  • LOC100347407
  • c-raf
  • Raf1
  • ZHX2
  • Zhx2
Epitop
AA 25-60
80
74
47
47
46
43
41
34
34
28
27
23
21
19
18
18
15
15
15
15
15
14
7
7
7
6
6
4
4
4
3
3
3
3
3
3
3
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Reaktivität
Human, Maus, Ratte (Rattus)
834
411
386
25
23
16
16
13
8
8
8
5
4
4
2
2
1
1
1
Wirt
Kaninchen
779
63
12
6
2
Klonalität
Polyklonal
Konjugat
Dieser RAF1 Antikörper ist unkonjugiert
45
45
44
30
29
29
14
14
12
12
12
12
12
12
12
12
12
2
2
2
2
2
2
2
2
2
1
1
Applikation
Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
512
365
221
172
155
103
52
49
20
18
18
7
2
2
2
1
Optionen
Hersteller
Anmelden zum Anzeigen
Hersteller Produkt- Nr.
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'Independent Validation' Siegel
Chargennummer 130909
Validierte Anwendung Immunohistochemistry
Positivkontrolle Human colon villus epithelial cells
Negativkontrolle Human liver
Bewertung Raf1 exhibits uniformly low expression across all human tissues; higher expression found in colon, lower in liver. Tissue-specific expression is detected in colon, with expected low level expression in liver.
Primärantikörper
Isotypkontrolle
Sekundärantikörper
Kontrollen
  • Positive control: Human colon tissue stained with antibody
  • Negative control: Human liver tissue stained with antibody
  • Isotype control: Human colon tissue stained with isotype control
  • Secondary only control: Human colon tissue stained with secondary antibody only
Protokoll
  • Immunohistochemistry was performed on a Ventana NEXes automated platform; instrument manufacturer specific reagents are italicized.
  • 1. Slides were preheated in convection oven at 60°C for 30 min
  • 2. Deparaffinization procedure: - 3 changes of Xylene, 5 min each - 3 changes of 100% Ethanol, 3 min each - 3 changes of 95% Ethanol, 3 min each - Rinsed in distilled water, 3 changes
  • 3. Heat retrieval procedure - Slides retrieved in 10.0 mM Citrate, pH6.0 in a 1000W microwave oven (~100°C) for 15 min. - Slides were allowed to cool (in citrate) for 30 min. - Slides were washed x 3 in Distilled water
  • 4. NEXes instrument procedure, iView DAB paraffin protocol (*abridged*): - Slide chamber warmed to 37°C
  • 5. Slides rinsed with *reaction buffer* x3
  • 6. *iView Inhibitor (H2O2)* applied and incubated for 4 min
  • 7. Slides rinsed with *reaction buffer*
  • 8. Antibody Application - Primary antibody diluted 1:250 in PBS (100 microliter applied/slide) - Ventana Isotype control applied neat - Slides Incubated overnight at room temperature (~12 hours ~25°C)
  • 9. Slides rinsed with *reaction buffer* x3
  • 10. *iView Biotinylated IgG* applied and incubated for 8 min
  • 11. Slides rinsed with *reaction buffer*
  • 14. *iView Streptavidin-Horseradish Peroxidase* applied and incubated for 8 min
  • 15. Slides rinsed with *reaction buffer*
  • 16. *iView DAB/H2O2* applied and incubated for 8 min
  • 17. Slides rinsed with *reaction buffer*
  • 18. *iView Copper* applied and incubated for 4 min
  • 19. Slides rinsed with *reaction buffer*
  • 20. Slides washed in Dawn Detergent/tap water
  • 21. Counterstain Procedure - Hematoxylin (Leica 560 MX) 30 sec - Slides washed in tap water, 1 min - Decolorized (10% Acetic Acid in 70% ethanol), 1 min - Slides washed in tap water, 1 min - Bluing (Austin Clear Ammonia), 1 min - Slides washed in tap water, 1 min
  • 22. Dehydration/coverslipping procedure: - 3 changes of 95% Ethanol, 3 min each - 3 changes of 100% Ethanol, 3 min each - 3 changes of Xylene, 5 min each - Mounted with Permount
  • 23. Imaging: Leica SCN 400F Whole Slide Scanner with Digital Image Hub and Leica Slidepath software
Anmerkungen
  • Deviations from protocol/procedure supplied by manufacturer:
  • Step 1: Heated tissue 60°C for 30 minutes; manufacturer heats for 45 minutes.
  • Step 2: No ethanol wash was performed during deparaffinization; manufacturer includes 1 wash of 80% ethanol for 3 minutes.
  • Step 3.1: Slides were heated for 15 minutes; manufacturer provides a range of 15-20 minutes.
  • Step 3.2: Slides were cooled for 30 minutes; manufacturer cools for 20 minutes.
  • Step 4: Italicized reagents and incubation time are fixed instrument parameters.
  • Step 5: Secondary species-specific serum block not used; manufacturer blocks with 5% normal goat serum for 2 hours.
  • Step 8.1: Antibody diluted in PBS at 1:250; manufacture did not recommend diluent or dilution.
  • Step 8.2.1: Primary antibody incubated at room temperature overnight; manufacturer incubates overnight 4°C with agitation.
Validierungsbilder
Immunohistochemistry validation image for anti-V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1 (RAF1) (AA 25-60) antibody (ABIN733208) Figure 1: Human colon tissue stained with anti-RAF1 (brown) and counterstained with h...
Immunohistochemistry validation image for anti-V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1 (RAF1) (AA 25-60) antibody (ABIN733208) Figure 2: Human liver tissue stained with anti-RAF1 (brown) and counterstained with h...
Immunohistochemistry validation image for anti-V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1 (RAF1) (AA 25-60) antibody (ABIN733208) Figure 3: Human colon tissue stained with isotype control antibody (brown) and counte...
Immunohistochemistry validation image for anti-V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1 (RAF1) (AA 25-60) antibody (ABIN733208) Figure 4: Human colon tissue stained with secondary atnibody only (brown) and counter...
Immunogen KLH conjugated synthetic peptide derived from human c Raf
Isotyp IgG
Reinigung Purified by Protein A.
Andere Bezeichnung C-Raf/Raf1 (RAF1 Antibody Abstract)
Hintergrund

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. Phosphorylates PPP1R12A resulting in inhibition of the phosphatase activity. Phosphorylates TNNT2/cardiac muscle troponin T. Can promote NF-kB activation and inhibit signal transducers involved in motility (ROCK2), apoptosis (MAP3K5/ASK1 and STK3/MST2), proliferation and angiogenesis (RB1). Can protect cells from apoptosis also by translocating to the mitochondria where it binds BCL2 and displaces BAD/Bcl2-antagonist of cell death. Regulates Rho signaling and migration, and is required for normal wound healing. Plays a role in the oncogenic transformation of epithelial cells via repression of the TJ protein, occludin (OCLN) by inducing the up-regulation of a transcriptional repressor SNAI2/SLUG, which induces down-regulation of OCLN. Restricts caspase activation in response to selected stimuli, notably Fas stimulation, pathogen-mediated macrophage apoptosis, and erythroid differentiation.

Subcellular location: Cytoplasm, Nucleus, Mitochondrion, Cell membrane

Synonyms: NS5, CRAF, Raf-1, c-Raf, CMD1NN, RAF proto-oncogene serine/threonine-protein kinase, Proto-oncogene c-RAF, RAF1, RAF

Gen-ID 5894
UniProt P04049
Pathways MAPK Signalweg, RTK Signalweg, Fc-epsilon Rezeptor Signalübertragung, Neurotrophin Signalübertragung, cAMP Metabolic Process, Stem Cell Maintenance, Hepatitis C, Autophagie, Signaling of Hepatocyte Growth Factor Receptor
Applikationshinweise WB 1:100-1000
IHC-P 1:100-500
IF(IHC-P) 1:50-200
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 1 μg/μL
Buffer Aqueous buffered solution containing 1 % BSA, 50 % glycerol and 0.09 % sodium azide.
Konservierungsmittel Sodium azide
Vorsichtsmaßnahmen This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
Lagerung -20 °C
Informationen zur Lagerung Store at -20°C
Haltbarkeit 12 months
Bilder des Herstellers
Immunohistochemistry (IHC) image for anti-V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1 (RAF1) (AA 25-60) antibody (ABIN733208) Formalin-fixed and paraffin embedded human breast cancer labeled with Anti-c-RafRaf1 ...
Immunohistochemistry (IHC) image for anti-V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1 (RAF1) (AA 25-60) antibody (ABIN733208) Images provided the Independent Validation Program (badge number 29651)Formalin-fixed...
Produkt verwendet in: Wang, Fu, Jiang, Yu, Wang, Han, Liu, Wang: "Bone marrow mononuclear cell transplantation promotes therapeutic angiogenesis via upregulation of the VEGF-VEGFR2 signaling pathway in a rat model of vascular dementia." in: Behavioural brain research, Vol. 265, pp. 171-80, 2014 (PubMed).

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