Overview

  • Product name
  • Description
    Goat polyclonal to RKIP
  • Host species
    Goat
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse, Cow, Dog
  • Immunogen

    Synthetic peptide corresponding to Human RKIP aa 174-186 (C terminal).
    Sequence:

    C-DYVPKLYEQLSGK


    (Peptide available as ab22921)

  • Positive control
    • WB: A549 and HepG2 cells lysates, human liver tissue lysate. IHC-P: Human brain (cortex) tissue.
  • General notes
    GenBank Accession Number – NP_002558.

Properties

Applications

Our Abpromise guarantee covers the use of ab2634 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 Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 20-22 kDa (predicted molecular weight: 22 kDa).Can be blocked with Human RKIP peptide (ab22921).

Target

  • Function
    Binds ATP, opioids and phosphatidylethanolamine. Has lower affinity for phosphatidylinositol and phosphatidylcholine. Serine protease inhibitor which inhibits thrombin, neuropsin and chymotrypsin but not trypsin, tissue type plasminogen activator and elastase.
    HCNP may be involved in the function of the presynaptic cholinergic neurons of the central nervous system. HCNP increases the production of choline acetyltransferase but not acetylcholinesterase. Seems to be mediated by a specific receptor.
  • Sequence similarities
    Belongs to the phosphatidylethanolamine-binding protein family.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • Epididymis luminal protein 210 antibody
    • Epididymis secretory protein Li 34 antibody
    • Epididymis secretory protein Li 96 antibody
    • HCNP antibody
    • HCNPpp antibody
    • HEL 210 antibody
    • HEL S 34 antibody
    • HEL S 96 antibody
    • Hippocampal cholinergic neurostimulating peptide antibody
    • Neuropolypeptide h3 antibody
    • PBP antibody
    • PEBP antibody
    • PEBP 1 antibody
    • PEBP-1 antibody
    • Pebp1 antibody
    • PEBP1_HUMAN antibody
    • Phosphatidylethanolamine binding protein antibody
    • Phosphatidylethanolamine binding protein 1 antibody
    • Prostatic binding protein antibody
    • Prostatic-binding protein antibody
    • R kip antibody
    • Raf kinase inhibitor protein antibody
    • Raf kinase inhibitory protein antibody
    • RKIP antibody
    see all

Images

  • Anti-RKIP antibody (ab2634) at 1 µg/ml + A549 lysate at 35 µg

    Predicted band size: 22 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cortex tissue labelling RKIP with ab2634 at 5µg/ml. Heat induced antigen retrieval was performed with citrate buffer pH 6. HRP-staining.

  • Negative control Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cortex tissue using no primary antibody.

References

This product has been referenced in:
  • Chen Y  et al. The expression, induction and pharmacological activity of CYP1A2 are post-transcriptionally regulated by microRNA hsa-miR-132-5p. Biochem Pharmacol 145:178-191 (2017). WB . Read more (PubMed: 28822783) »
  • Fougner SL  et al. Low levels of raf kinase inhibitory protein in growth hormone-secreting pituitary adenomas correlate with poor response to octreotide treatment. J Clin Endocrinol Metab 93:1211-6 (2008). WB ; Human . Read more (PubMed: 18230656) »
See all 3 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Abreviews
Application
Western blot
Sample
Rat Tissue lysate - whole (Hippocampus)
Loading amount
50 µg
Specification
Hippocampus
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Dr. Martin Maurer

Verified customer

Submitted Oct 31 2006

Answer

I'm very sorry to hear you are experiencing problems with ab2634. Thank you for taking the time to provide your protocol details and image, those helped me a lot understand your problem. The fact that you detect the His-PBP protein with a His antibody suggests that potentially the protein levels in the first sample is high and should be detectable with ab2634. However, can you please clarify if the recombinant protein is the whole protein or a fragment? The antibody is designed to recognise aa 174-188 and therefore if your protein does not contain this fragment the antibody cannot recognise it.Is the protein originally from the human form? If you could run in parallel a positive control of A549, HepG2 or Human Liver lysate, these are guaranteed to work with ab2634 and will also tell you if your other samples of SH-SY5Y human neuroblastoma cells have high levels of PBP protein. I am a little confused about the title of your image " digestion by calpain". Have you incubated your cells prior to extraction of the protein? Calpain digestion may be damaging your samples. The antibody has been tested against reduced samples so the high bands may also be due to the fact you use non-reduced conditions. The absence of a specific band can also be due to the fact that the antibody has not bound for sufficient amount of time to the membrane and the non specific bands could also be due to the blocking agent. To test this hypothesis can I please suggest blocking the membrane in 5% BSA in TBST for 1hr, rinsing lightly the membrane in TBST and incubating the antibody in TBST only (the Tween percentage should be 0.1%)overnight at 4C. You may be able to dilute the antibody more as it will be incubated overnight, giving you a slow but targeted binding of the antibody. I hope these suggestions will help you, please do not hesitate to contact me again if I can be of further help,

Read More

Answer

My apologies for the delay, I have just received the following information from the source of those two antibodies: -ab2634 Before loading samples had 5% (v/v) beta-mercaptoethanol was added and were boiled for 3 minutes. -ab16050 This antibody was made from recombinant protein of 206 amino acids. The originator of that antibody has the recombinant protein in stock but doesn't know if it can be used as blocker so I would not recommend it for blocking/competitive studies. I hope this information helps, please do not hesitate to contact us again if you need further information,

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