Overview

  • Product name

  • Description

    Rabbit polyclonal to IB-1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse IB-1 (N terminal).

  • Positive control

    • Mouse Intestine. Rat kidney. Jurkat whole cell lysate (ab7899).
  • General notes

    Protein previously labeled as JIP1.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.02% Thimerosal (merthiolate)
    Constituents: PBS, 50% Glycerol, 0.1% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab24449 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 5 µg/ml.
WB Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 78 kDa (predicted molecular weight: 78 kDa). A band of 110 kDa can also be observed in human samples.

Target

  • Relevance

    Function: The JNK-interacting protein (JIP) group of scaffold proteins selectively mediates JNK signaling by aggregating specific components of the MAPK cascade to form a functional JNK signaling module. Required for JNK activation in response to excitotoxic stress. Cytoplasmic MAPK8IP1 causes inhibition of JNK-regulated activity by retaining JNK in the cytoplasm and inhibiting JNK phosphorylation of c-Jun. May also participate in ApoER2-specific reelin signaling. Directly, or indirectly, regulates GLUT2 gene expression and beta-cell function. Appears to have a role in cell signaling in mature and developing nerve terminals. May function as a regulator of vesicle transport, through interactions with the JNK-signaling components and motor proteins (By similarity). Functions as an anti-apoptotic protein and whose level seems to influence the beta-cell death or survival response. Tissue specificity: Highly expressed in brain. Expressed in neurons, localizing to neurite tips in differentiating cells. Also expressed in the pancreas, testis and prostate. Low levels in heart, ovary and small intestine. Decreased levels in pancreatic beta cells sensitize cells to IL-1-beta-induced apoptosis. Disease: Diabetes mellitus, non-insulin-dependent Similarity: Belongs to the JIP scaffold family. Contains 1 PID domain. Contains 1 SH3 domain. Domain: The destruction boxes (D-box) may act as recognition signals for degradation via the ubiquitin-proteasome pathway. A minimal inhibitory domain prevents pancreatic beta cell apoptosis in vitro, and prevents activation of c-jun by MAPK8, MAPK9 and MAPK10. The SH3 domain mediates homodimerization. PTM: Phosphorylated by MAPK8, MAPK9 and MAPK10. Phosphorylation on Thr-103 is also necessary for the dissociation and activation of MAP3K12. Phosphorylated by isoform 1 and isoform 2 of VRK2. Hyperphosphorylated during mitosis following activation of stress-activated and MAP kinases. Ubiquitinated. Two preliminary events are required to prime for ubiquitination; phosphorylation and an increased in intracellular calcium concentration. Then, the calcium influx initiates ubiquitination and degradation by the ubiquitin-proteasome pathway.
  • Cellular localization

    Cytoplasmic. Accumulates in cell surface projections. Under certain stress conditions, translocates to the perinuclear region of neurons. In insulin-secreting cells, detected in both the cytoplasm and nucleus
  • Database links

  • Alternative names

    • C jun amino terminal kinase interacting protein 1 antibody
    • C-jun-amino-terminal kinase-interacting protein 1 antibody
    • IB 1 antibody
    • IB-1 antibody
    • IB1 antibody
    • Islet brain 1 antibody
    • Islet-brain 1 antibody
    • JIP 1 antibody
    • JIP-1 antibody
    • JIP1_HUMAN antibody
    • JNK interacting protein 1 antibody
    • JNK MAP kinase scaffold protein 1 antibody
    • JNK-interacting protein 1 antibody
    • MAPK8IP 1 antibody
    • MAPK8IP1 antibody
    • MAPK8IP1 mitogen-activated protein kinase 8 interacting protein 1 antibody
    • Mitogen activated protein kinase 8 interacting protein 1 antibody
    • Mitogen-activated protein kinase 8-interacting protein 1 antibody
    • PRKM8 antibody
    • PRKM8 interacting protein antibody
    • PRKM8IP antibody
    see all

Images

  • All lanes : Anti-IB-1 antibody (ab24449)

    Lane 1 : Rat kidney
    Lane 2 : Mouse Intestine
    Lane 3 : Jurkat Cell lysate

    Predicted band size: 78 kDa
    Observed band size: 78 kDa
    Additional bands at: 110 kDa (possible isoform)

  • ICC/IF image of ab24449 stained HepG2 cells. The cells were 4% formaldehyde 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 (ab24449, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 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.

References

This product has been referenced in:

  • Sherman MA  et al. Soluble Conformers of Aß and Tau Alter Selective Proteins Governing Axonal Transport. J Neurosci 36:9647-58 (2016). Read more (PubMed: 27629715) »
See 1 Publication for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting us again.
Please contact my colleges from the financial department, as I can raise a credit note, but everything else is in their hands.

It may be useful to contact the Apotheke as well to inform that this credit note is exclusively for your group and not to be used for paying outstanding invoices (we know from experience that many university purchasing agent do this).
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Use our products? Submit an Abreview. Earn rewards! https://www.abcam.com/abreviews

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Answer

Thank you for contacting us.
Your credit note ID is 22405.
I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:
(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.
Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.
To specifically receive a refund please contact Tel# 030 896 779 154.
The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.
I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

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Question

Dear Anja

Attached you will find the information you required. The images of GFP-JIP1 transfected neurons stained with anti-JIP1 show almost no co-localization.

1) Abcam product code ab 24449

2) Abcam order reference number or product batch number
Purchase order number: 30072012
3) Description of the problem the staining is unspecific, looks like tubulin, but is in rat hippocampal neurons not localized to the axon tip (like expected and observed with JIP1-GFP. Shows also no signif. Co-localization with JIP1-GFP!
4) Sample preparation: primary neurons prepared to common protocols and seeded on poly-ornithin coated coverslips like usual. cells fixed with 4% PFA-succrose, blocked with NGS
Species rat,
Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other: primary hippocampal neurons, fixed with PFA(4%) -Succrose
Sample preparation standard protocol for primary neuron culture
Positive control JIP1-GFP transfected into the neuron by AMAXA electroporation
Negative control secondary ab alone (alexa 488/594, Invitrogen)
5) Fixation step
Yes/No
If yes: Fixative agent and concentration PFA 4% - succrose
Fixation time 18 min
Fixation temperature RT
6) Antigen retrieval method no, not recommended for this cells
7) Permeabilization method:
Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers? Yes
Permeabilizing agent and concentration:0.1% Triton-X, 0.1% Na-Citrate in PBS for 4 min at RT

8) Blocking agent (eg BSA, serum…): 10% NGS in PBS
Concentration
Blocking time 1h
Blocking temperature RT
9) Endogenous peroxidases blocked? no
Endogenous biotins blocked? no
10) Primary antibody (If more than one was used, describe in “additional notes”) :
Concentration or dilution 1:40 (tried also 1:100; 1:25)
Diluent buffer 5% NGS in PBS
Incubation time ON at 4°C
11) Secondary antibody:
Species: goat
Reacts against: rabbit
Concentration or dilution 1:1000
Diluent buffer PBS
Incubation time 90 min
Fluorochrome or enzyme conjugate alexa 488/ alexa 594
12) Washing after primary and secondary antibodies:
Buffer pbs
Number of washes 4
13) Detection method fluorescence
14) How many times have you run this staining? 2x
Do you obtain the same results every time? yes
What steps have you altered to try and optimize the use of this antibody? Changed concentrations; added tween-20; included JIP1-GFP transfection as positive control
Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem. Please find attached a pic with neurons transfected with JIP1-GFP (green), staind with JIP1 ab (ab24449) +Alexa 594 (red)

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Answer

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.
As Anja is in her holidays, i will deal with your case.
The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.
Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial. In addition, Anja had reviewed the images from the lab on rat neuron cells and the results fit your discribtion of the localization.
I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation.
Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Question
Answer

Thank you for contacting Abcam.

I am sorry that this product did not perform correctly for you. As we had spoken about, if this product has been purchased within the last six months it will be covered by our Abpromise guarantee and I would be able to offer you a free of charge replacement for this. When you have decided which product you would like for that replacement please let me know either by phone or by email. In order to process the replacement we will need the Abcam order confirmation number as well.

Please let me know if you have any questions or concerns. I look forward to your reply.

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Answer

Thank you for your enquiry. To our knowledge, the antibody has yet to be tested in immunoprecipitation. All tested applications are specified on Abcam product datasheets. If you decide to go ahead and purchase this product, we encourage you to submit an Abreview via the online product datasheet. We always encourage customers to send their results back to us, whether positive or negative, and make all product information available to researchers. To find out more about our Abreview system, please see the following URL: https://www.abcam.com/abreviews I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

I'm afraid I have been quite unsuccessful at finding out the information you needed. The source of the antibody did not wish to divulge to me the exact sequence and I was only able to find out that it was the N terminal part of the protein which was used. I'm sorry I couldn't help you more on this occasion,

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