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

Details for Product anti-RAF1 Antibody No. ABIN733208, Supplier: Log in to see
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
Alternatives
anti-Rat (Rattus) RAF1 antibody for Dot Blot
Epitope
AA 25-60
73
72
47
40
39
37
28
28
28
27
22
22
22
20
19
18
17
15
15
15
15
14
14
14
10
8
8
7
6
6
6
6
6
4
3
3
3
3
3
3
3
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
Reactivity
Human, Mouse (Murine), Rat (Rattus)
808
410
385
25
23
16
16
13
8
8
8
5
4
4
2
2
1
1
1
Host
Rabbit
769
46
12
6
2
Clonality
Polyclonal
Conjugate
This RAF1 antibody is un-conjugated
44
44
43
29
29
29
13
13
12
12
12
12
12
12
12
12
12
1
1
1
1
1
1
1
1
1
Application
Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
489
348
202
172
155
103
50
34
19
18
17
7
2
2
2
1
Options
Supplier
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Available images

'Independent Validation' Badge
Lot Number 130909
Method validated Immunohistochemistry
Positive Control Human colon villus epithelial cells
Negative Control Human liver
Notes 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.
Primary Antibody
  • Antigen: V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1 (RAF1)
  • Catalog number: ABIN733208
  • Supplier: Log in to see
  • Supplier catalog number: Log in to see
  • Lot number: 130909
Isotype Control Antibody
Secondary Antibody
Controls
  • 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
Protocol
  • 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
Experimental Notes
  • 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.
Validation Images
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
Isotype IgG
Purification Purified by Protein A.
Alternative Name C-Raf/Raf1 (RAF1 Antibody Abstract)
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. 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

Gene ID 5894
UniProt P04049
Pathways MAPK Signaling, RTK Signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, cAMP Metabolic Process, Stem Cell Maintenance, Hepatitis C, Autophagy, Signaling of Hepatocyte Growth Factor Receptor
Application Notes WB 1:100-1000
IHC-P 1:100-500
IF(IHC-P) 1:50-200
Restrictions For Research Use only
Format Liquid
Concentration 1 μg/μL
Buffer Aqueous buffered solution containing 1 % BSA, 50 % glycerol and 0.09 % sodium azide.
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
Storage -20 °C
Storage Comment Store at -20°C
Expiry Date 12 months
Supplier Images
Immunohistochemistry (IHC) image for anti-RAF1 antibody (V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1) (AA 25-60) (ABIN733208) Formalin-fixed and paraffin embedded human breast cancer labeled with Anti-c-RafRaf1 ...
Immunohistochemistry (IHC) image for anti-RAF1 antibody (V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1) (AA 25-60) (ABIN733208) Images provided the Independent Validation Program (badge number 29651)Formalin-fixed...
Product cited 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).