Overview

  • Product name

  • Description

    Rabbit polyclonal to CABLES1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Zebrafish
  • Immunogen

    Synthetic peptide corresponding to Human CABLES1 aa 600 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab87824)

  • Positive control

    • This antibody gave a positive signal in Human Brain and Skeletal Muscle Tissue Lysates, and in the following whole cell lysates: SHSY-5Y; SK N SH; SK N BE; HeLa.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab75535 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 68 kDa (predicted molecular weight: 68 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    Cyclin-dependent kinase binding protein. Enhances cyclin-dependent kinase tyrosine phosphorylation by nonreceptor tyrosine kinases, such as that of CDK5 by activated ABL1, which leads to increased CDK5 activity and is critical for neuronal development, and that of CDK2 by WEE1, which leads to decreased CDK2 activity and growth inhibition. Positively affects neuronal outgrowth. Plays a role as a regulator for p53/p73-induced cell death.
  • Tissue specificity

    Expressed in breast, pancreas, colon, head and neck (at protein level). Strongly decreased in more than half of cases of atypical endometrial hyperplasia and in more than 90% of endometrial cancers.
  • Sequence similarities

    Belongs to the cyclin family.
  • Developmental stage

    Expression in the endometrial epithelium fluctuates during the menstrual cycle, being greater during the secretory phase when compared with the proliferative phase.
  • Post-translational
    modifications

    Phosphorylated on Ser-313 by CCNE1/CDK3. Phosphorylated on serine/threonine residues by CDK5 and on tyrosine residues by ABL1. Also phosphorylated in vitro by CCNA1/CDK2, CCNE1/CDK2, CCNA1/CDK3 and CCNE1/CDK3.
  • Cellular localization

    Nucleus. Cytoplasm. Located in the cell body and proximal region of the developing axonal shaft of immature neurons. Located in axonal growth cone, but not in the distal part of the axon shaft or in dendritic growth cone of mature neurons.
  • Information by UniProt
  • Database links

  • Alternative names

    • CABL1_HUMAN antibody
    • CABLES antibody
    • Cables1 antibody
    • Cdk5 and Abl enzyme substrate 1 antibody
    • CDK5 and ABL1 enzyme substrate 1 antibody
    • FLJ35924 antibody
    • HsT2563 antibody
    • IK3 1 antibody
    • Ik3-1 antibody
    • Interactor with CDK3 1 antibody
    see all

Images

  • IHC image of CABLES1 staining in Human Breast Carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab5535, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • All lanes : Anti-CABLES1 antibody (ab75535) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 3 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 4 : SK N SH (Human neuroblastoma) Whole Cell Lysate
    Lane 5 : SK N BE (Human neuroblastoma) Whole Cell Lysate
    Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 68 kDa
    Observed band size: 68 kDa
    Additional bands at: 42 kDa (possible isoform)



    This antibody is predicted to cross react with both isoform 1 (68 kDa) and isoform 2 (42 kDa) of CABLES1. We believe that the band observed at 42 kDa corresponds to isoform 2.
  • ICC/IF image of ab75535 stained MCF-7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75535, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HEK293, HepG2 cells at 1µg/ml, and in 100% Methanol fixed (5 min) HeLa, HEK293, HepG2, and MCF-7 cells at 5µg/ml.

References

This product has been referenced in:

  • Hernández-Ramírez LC  et al. Loss-of-function mutations in theCABLES1gene are a novel cause of Cushing's disease. Endocr Relat Cancer 24:379-392 (2017). Read more (PubMed: 28533356) »
  • Roussel-Gervais A  et al. The Cables1 Gene in Glucocorticoid Regulation of Pituitary Corticotrope Growth and Cushing Disease. J Clin Endocrinol Metab 101:513-22 (2016). Read more (PubMed: 26695862) »
See all 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1048177.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

PS: As discussed please send the images for further investiagtion.

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Answer

Thank you for contacting us with your question. We don't have a specific protocol for using this antibody with skin tissues, but we would recommend following our general IHC-P protocol which I have attached to this email. The only specific recommendation that we have for this antibody is to use heat mediated antigen retrieval prior to staining. The optimal concentration of antibody is dependent on the particular sample, but we recommend starting around 1 ug/mL of ab75535. In order to calculate the exact total volume of antibody you need, you can first determine the optimal dilution of antibody for your skin samples by testing a few different dilutions and comparing the results. That being said, we can estimate how much antibody you will need. Ab75535 is provided as 100 ug of 1 mg/mL solution. So, if 1 ug/mL is indeed the optimal dilution for your samples, you will have 100 mL of working solution. For 1 slide, you'll use around 100 uL of working solution, so in this case you would have enough working solution for 1000 slides. You may be able to fit more than one sample on one slide depending on the diameter of the sections. So, one vial of ab75535 is probably enough antibody to stain all of your samples. I hope this information will be useful, but please let me know if there is anything else you need and I'll be happy to help.

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