Directly conjugated antibodies for immunofluorescence

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Overview

  • Product nameAnti-Raf1 antibody [Y198]
    See all Raf1 primary antibodies
  • Description
    Rabbit monoclonal [Y198] to Raf1
  • Tested applicationsIHC-P, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive)

  • Positive control
    • NIH 3T3 cell lysate and human colon carcinoma
  • General notes

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab32025 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/400.

For unpurified, use 1/30.

ICC/IF 1/400.

For unpurified, use 1/30.

WB 1/1000 - 1/10000. Detects a band of approximately 74 kDa (predicted molecular weight: 73 kDa).

For unpurified, use 1/200.

  • Application notesIs unsuitable for Flow Cyt or IP.
  • Target

    • FunctionInvolved in the transduction of mitogenic signals from the cell membrane to the nucleus. Part of the Ras-dependent signaling pathway from receptors to the nucleus. Protects cells from apoptosis mediated by STK3.
    • Tissue specificityIn skeletal muscle, isoform 1 is more abundant than isoform 2.
    • Involvement in diseaseDefects in RAF1 are the cause of Noonan syndrome type 5 (NS5) [MIM:611553]. Noonan syndrome (NS) is a disorder characterized by dysmorphic facial features, short stature, hypertelorism, cardiac anomalies, deafness, motor delay, and a bleeding diathesis. It is a genetically heterogeneous and relatively common syndrome, with an estimated incidence of 1 in 1000-2500 live births.
      Defects in RAF1 are the cause of LEOPARD syndrome type 2 (LEOPARD2) [MIM:611554]. LEOPARD syndrome is an autosomal dominant disorder allelic with Noonan syndrome. The acronym LEOPARD stands for lentigines, electrocardiographic conduction abnormalities, ocular hypertelorism, pulmonic stenosis, abnormalities of genitalia, retardation of growth, and deafness.
    • Sequence similaritiesBelongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily.
      Contains 1 phorbol-ester/DAG-type zinc finger.
      Contains 1 protein kinase domain.
      Contains 1 RBD (Ras-binding) domain.
    • Post-translational
      modifications
      Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylation at Thr-269 increases its kinase activity. Phosphorylation at Ser-259 induces the interaction with YWHAZ and inactivates kinase activity. Dephosphorylation of Ser-259 by the complex containing protein phosphatase 1, SHOC2 and M-Ras/MRAS relieves inactivation, leading to stimulate RAF1 activity.
    • Cellular localizationCytoplasm. Cell membrane. Colocalizes with RGS14 and BRAF in both the cytoplasm and membranes.
    • Information by UniProt
    • Database links
    • Alternative names
      • c Raf antibody
      • C-raf antibody
      • C-Raf proto-oncogene, serine/threonine kinase antibody
      • CMD1NN antibody
      • Craf 1 transforming gene antibody
      • Craf 1 transforming gene antibody
      • cRaf antibody
      • cRaf antibody
      • Craf1 transforming gene antibody
      • EC 2.7.11.1 antibody
      • kinase Raf1 antibody
      • Murine sarcoma 3611 oncogene 1 antibody
      • Murine sarcoma 3611 oncogene 1 antibody
      • NS5 antibody
      • Oncogene MIL antibody
      • Oncogene RAF1 antibody
      • OTTHUMP00000160218 antibody
      • OTTHUMP00000207813 antibody
      • OTTHUMP00000209389 antibody
      • Protein kinase raf 1 antibody
      • Proto-oncogene c-RAF antibody
      • Raf 1 antibody
      • Raf 1 proto oncogene serine/threonine kinase antibody
      • RAF antibody
      • Raf proto oncogene serine/threonine protein kinase antibody
      • RAF proto-oncogene serine/threonine-protein kinase antibody
      • RAF-1 antibody
      • RAF1 antibody
      • RAF1_HUMAN antibody
      • Similar to murine leukemia viral (V-raf-1) oncogene homolog 1 antibody
      • TRANSFORMING REPLICATION-DEFECTIVE MURINE RETROVIRUS 3611-MSV antibody
      • v raf 1 murine leukemia viral oncogene homolog 1 antibody
      • v-raf murine sarcoma viral oncogene homolog 1 antibody
      • vraf1 murine leukemia viral oncogene homolog 1 antibody
      see all

    Anti-Raf1 antibody [Y198] images

    • Anti-Raf1 antibody [Y198] (ab32025) at 1/2000 dilution (purified) + NIH/3T3 cell lysate at 10 µg

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 73 kDa
      Observed band size : 74 kDa (why is the actual band size different from the predicted?)

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunohistochemical staining of paraffin embedded human liver with purified ab32025 at a working dilution of 1 in 200. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0

    • Immunofluorescence staining of MCF7 cells with purified ab32025 at a working dilution of 1 in 200, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 555 goat anti rabbit, used at a dilution of 1 in 200. The cells were fixed in 4% PFA.

    • Anti-Raf1 antibody [Y198] (ab32025) at 1/200 dilution (unpurified) + NIH/3T3 cell lysate at 10 µg

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 73 kDa
      Observed band size : 74 kDa (why is the actual band size different from the predicted?)

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunohistochemical staining of paraffin embedded human liver with unpurified ab32025 at a working dilution of 1 in 30. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0

    • Immunofluorescence staining of MCF7 cells with unpurified ab32025 at a working dilution of 1 in 30, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 555 goat anti rabbit, used at a dilution of 1 in 200. The cells were fixed in 4% PFA.

    • Anti-Raf1 antibody [Y198] (ab32025) at 1/500 dilution (unpurified) + NIH 3T3 cell lysate

      Predicted band size : 73 kDa
      Observed band size : 74 kDa (why is the actual band size different from the predicted?)
    • Immunohistochemical staining of human Raf1 in paraffin embedded colon carcinoma with unpurified ab32025, at a 1/100 dilution.

      ab32025, at a 1/100 dilution, staining human Raf1 in colon carcinoma by Immunohistochemistry, Paraffin embedded tissue

    References for Anti-Raf1 antibody [Y198] (ab32025)

    This product has been referenced in:
    • Mehta S  et al. Predictive factors in patients with advanced and metastatic squamous cell carcinoma of the head and neck: a study based on SWOG protocol S0420. Oncol Rep 29:2095-100 (2013). Read more (PubMed: 23563900) »
    • Miller JP  et al. A genome-scale RNA-interference screen identifies RRAS signaling as a pathologic feature of Huntington's disease. PLoS Genet 8:e1003042 (2012). WB ; Human . Read more (PubMed: 23209424) »

    See all 7 Publications for this product

    Product Wall

    Thank you for your reply.
    According to the attached slide, we detected a signal using NIH/3T3 untreated and treated with PDGF. Normally, for a phospho specific antibody, no signal is seen without PDGF treatment. However, a pan antibody recognizes b...

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    Thank you for contacting Abcam yesterday.

    I have been in contact with our lab and the antibody Raf-1 (ab32025) is a pan antibody, so it recognizes both unphosphorylated and phosphorylated protein. The antibody will bind to the phosphorylated f...

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