Overview

  • Product name

  • Description

    Rabbit polyclonal to EIF3F
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, ELISA, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Chimpanzee
    Predicted to work with: Cynomolgus monkey
  • Immunogen

    Synthetic peptide:

    ARVIGTLLGTVD

    , corresponding to amino acids 114-125 of Human EIF3F

  • Positive control

    • Human brain cerebellum tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab64177 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
WB 1/200 - 1/2000. Predicted molecular weight: 38 kDa.
IP Use at an assay dependent concentration.
ELISA 1/10000 - 1/40000.
IHC-P Use a concentration of 2.5 µg/ml.
ICC/IF Use at an assay dependent concentration.

Target

  • Function

    Component of the eukaryotic translation initiation factor 3 (eIF-3) complex, which is required for several steps in the initiation of protein synthesis. The eIF-3 complex associates with the 40S ribosome and facilitates the recruitment of eIF-1, eIF-1A, eIF-2:GTP:methionyl-tRNAi and eIF-5 to form the 43S preinitiation complex (43S PIC). The eIF-3 complex stimulates mRNA recruitment to the 43S PIC and scanning of the mRNA for AUG recognition. The eIF-3 complex is also required for disassembly and recycling of post-termination ribosomal complexes and subsequently prevents premature joining of the 40S and 60S ribosomal subunits prior to initiation.
    Deubiquitinates activated NOTCH1, promoting its nuclear import, thereby acting as a positive regulator of Notch signaling.
  • Sequence similarities

    Belongs to the eIF-3 subunit F family.
    Contains 1 MPN (JAB/Mov34) domain.
  • Domain

    The MPN domain mediates deubiquitinating activity.
  • Post-translational
    modifications

    Phosphorylation is enhanced upon serum stimulation. Phosphorylated during apoptosis by caspase-processed CDK11.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Database links

  • Alternative names

    • Deubiquitinating enzyme eIF3f antibody
    • eIF 3 epsilon antibody
    • eIF-3-epsilon antibody
    • eIF3 p47 antibody
    • eIF3 p47 subunit antibody
    • eIF3-epsilon antibody
    • eIF3-p47 antibody
    • eIF3f antibody
    • EIF3F_HUMAN antibody
    • EIF3S5 antibody
    • EIF3S5, formerly antibody
    • Eukaryotic translation initiation factor 3 subunit 5 antibody
    • Eukaryotic translation initiation factor 3 subunit 5, formerly antibody
    • Eukaryotic translation initiation factor 3 subunit F antibody
    • eukaryotic translation initiation factor 3, subunit 5 (epsilon, 47kD) antibody
    • eukaryotic translation initiation factor 3, subunit 5 epsilon, 47kDa antibody
    • eukaryotic translation initiation factor 3, subunit F antibody
    • translation initiation factor 3 47 kda subunit antibody
    see all

Images

  • ab64177 at 2.5µg/ml staining EIF3F in human brain cerebellum by Immunohistochemistry, Formalin-fixed, paraffin-embedded tissue.
  • ICC/IF image of ab64177 stained PC12 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 (ab64177, 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.

References

This product has been referenced in:

  • Parajuli P  et al. Twist1 Activation in Muscle Progenitor Cells Causes Muscle Loss Akin to Cancer Cachexia. Dev Cell 45:712-725.e6 (2018). Read more (PubMed: 29920276) »
  • Lang CH  et al. BCATm deficiency ameliorates endotoxin-induced decrease in muscle protein synthesis and improves survival in septic mice. Am J Physiol Regul Integr Comp Physiol 299:R935-44 (2010). Read more (PubMed: 20554928) »
See all 2 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you for confirming this information and for your help and cooperation with this case.

As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order.

Credit ID: #####


As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. Please refer to the credit ID number in any correspondence with the accounting department.

I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.

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Question

Product code: 64177
Lot number: GR30438-5

Inquiry: No signal at predicted size. Please check this.


1) Abcam product code :
ab64177

2) Abcam order reference number or product batch number :
GR30438-5,

2-1) storage temperature of antibody : -80℃

3) Description of the problem :
No signal

4) Sample preparation:
Type of sample and species(whole cell lysates, fraction, recombinant protein;and Human, mouse): Whole cell lysates , Human, SW480 cell line
Lysis buffer : RIPA lysis beffer
Protease inhibitors: 1x protesase inhibitor cocktail (sigma)
Phosphatase inhibitors: 1x phosphatase inhibitor cocktail (sigma)
Reducing agent: No
Boiling for ≥5 min? yes (10min)
Protein loaded ug/lane or cells/lane : 30ug/ lane
Positive control : si-GFP sample and FLAG-eIF3f over expression sample
Negative control : si-eIF3f sample

5) Percentage of gel : 12% SDS-PAGE
Type of membrane : PVDF membrane(Bio-rad)
Protein transfer verified : Ponceau S
Blocking agent and concentration : 5% Normal Horse Serum in 1x TBST
Blocking time : 1hr
Blocking temperature : Room Temperature

6) Primary antibody (If more than one was used, describe in “additional notes”) :
Concentration or dilution : 1:1000 and 1:500
Diluent buffer : 5% Normal Horse Serum in 1x TBST
Incubation time : 12hr
Incubation temperature: 4℃

7) Secondary antibody: peroxidase-conjugated Affinipure Donkey anti-Rabbit IgG(H+L) (711-035-152, Jackson ImmunoReserch)
Species: Donkey
Isotype: IgG
Reacts against:
Concentration or dilution : 1: 2000
Diluent buffer : 1x TBST
Incubation time : 1hr
Incubation temperature: Room Temperature
Fluorochrome or enzyme conjugate: HRP

8) Washing after primary and secondary antibodies:
Buffer : 1x TBST
Number of washes : each 3 times for 10min

9)Detection method : ECL solution method
10) How many times have you run this staining? 3 times
-Do you obtain the same results every time? 3 times
-Have you run a No Primary; control? no
-What steps have you altered to try and optimize the use of this antibody?
I used low to high titer for this antibody. (1:1000 ˜1:500)



Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

Our customer confirmed elF3f level through RT-PCR. But they didn't show any difference of elF3f level in endogenous, knock down modeled, over-expressed human sw480cell. also, they didn't show bands at predicted size.

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will provide us with vital information for our monitoring of product quality

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been succesful. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably recieved a bad vial.

I apologise for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation.

In addition, I can suggest you may like to consider including a no primary control in future experiments, toassess if the secondary antibody is binding non specifically.

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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