Overview

  • Product name

    Anti-EFEMP1/Fibulin-3 antibody
    See all EFEMP1/Fibulin-3 primary antibodies
  • Description

    Rabbit polyclonal to EFEMP1/Fibulin-3
  • Host species

    Rabbit
  • Specificity

    ab106429 is predicted not to cross-react with other Fibulin proteins.
  • Tested applications

    Suitable for: ICC/IF, WB, ICCmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human EFEMP1/Fibulin-3 (C terminal).
    Database link: Q12805

  • Positive control

    • HeLa cell lysate.
  • General notes

    Antibodies should not be exposed to prolonged high temperatures.

     This product was previously labelled as EFEMP1

     

Properties

Applications

Our Abpromise guarantee covers the use of ab106429 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 20 µg/ml.
WB Use a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 55 kDa.
ICC Use a concentration of 20 µg/ml.

Target

  • Function

    Binds EGFR, the EGF receptor, inducing EGFR autophosphorylation and the activation of downstream signaling pathways. May play a role in cell adhesion and migration. May function as a negative regulator of chondrocyte differentiation. In the olfactory epithelium, it may regulate glial cell migration, differentiation and the ability of glial cells to support neuronal neurite outgrowth.
  • Tissue specificity

    In the eye, associated with photoreceptor outer and inner segment regions, the nerve fiber layer, outer nuclear layer and inner and outer plexiform layers of the retina.
  • Involvement in disease

    Defects in EFEMP1 are a cause of Doyne honeycomb retinal dystrophy (DHRD) [MIM:126600]; also known as malattia leventinese (MLVT) (ML). DHRD is an autosomal dominant disease characterized by yellow-white deposits known as drusen that accumulate beneath the retinal pigment epithelium.
  • Sequence similarities

    Belongs to the fibulin family.
    Contains 6 EGF-like domains.
  • Cellular localization

    Secreted > extracellular space. Secreted > extracellular space > extracellular matrix. Localizes to the lamina propria underneath the olfactory epithelium.
  • Information by UniProt
  • Database links

  • Alternative names

    • DHRD antibody
    • Doyne honeycomb retinal degeneration antibody
    • DRAD antibody
    • EFEMP 1 antibody
    • EFEMP1 antibody
    • EGF containing fibulin like extracellular matrix protein 1 antibody
    • EGF containing fibulin like extracellular matrix protein 1 precursor antibody
    • EGF-containing fibulin-like extracellular matrix protein 1 antibody
    • Epidermal growth factor containing fibulin like extracellular matrix protein 1 antibody
    • Extracellular protein S1 5 antibody
    • Extracellular protein S1-5 antibody
    • FBLN 3 antibody
    • FBLN3 antibody
    • FBLN3_HUMAN antibody
    • FBNL antibody
    • FIBL 3 antibody
    • FIBL-3 antibody
    • FIBL3 antibody
    • Fibrillin like antibody
    • Fibrillin like protein antibody
    • Fibrillin-like protein antibody
    • FIBULIN 3 antibody
    • Fibulin-3 antibody
    • Fibulin3 antibody
    • FLJ35535 antibody
    • Heat shock 70 KD protein 1 antibody
    • MGC111353 antibody
    • MLVT antibody
    • MTLV antibody
    • S1 5 antibody
    • T16 protein antibody
    see all

Images

  • Lane 1 : Anti-EFEMP1/Fibulin-3 antibody (ab106429) at 0.5 µg/ml
    Lane 2 : Anti-EFEMP1/Fibulin-3 antibody (ab106429) at 1 µg/ml

    All lanes : HeLa cell lysate

    Lysates/proteins at 15 µg per lane.

    Predicted band size: 55 kDa

  • Immunohistochemical analysis of HeLa cells labelling EFEMP1/Fibulin-3 with ab106429 at 20 µg/ml. 

  • Immunofluorescence analysis of Hela cells labelling Fibulin 3 with ab106429 at 20 ug/ml.

References

This product has been referenced in:

  • Shao L  et al. Genomic expression profiles in cumulus cells derived from germinal vesicle and MII mouse oocytes. Reprod Fertil Dev N/A:N/A (2015). Read more (PubMed: 25989843) »
See 1 Publication 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 of ab106429. 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.

Read More

Question


My answers are after the questions,

1) Abcam product code ab106429

2) Abcam order reference number or product batch number: GR65345-2

3) Description of the problem: no bands or background

4) Sample preparation:

Type of sample (whole cell lysates, fraction, recombinant protein…): protein isolated from cell cultures (in the case of wb) and cryosections (in the case of IHC)

Lysis buffer: RLT buffer (Qiagen) for cell lysis

Protease inhibitors: Roche Tablets

Phosphatase inhibitors

Reducing agent: B-mercaptoethanol (tried also without this agent and the result was no bands)

Boiling for ≥5 min? yes/no boiling 5’ 95deg

Protein loaded ug/lane or cells/lane 20ug/lane

Positive control yes

Negative control yes (only secondary antibody)

5) Percentage of gel 4-12% Invitrogen precasted

Type of membrane PVDF

Protein transfer verified Yes, gel stained with Coomasie Blue after transference and it was fine

Blocking agent and concentration BSA 1% (IHC) Odyssey Blocking Buffer (WB)

Blocking time 1 hour

Blocking temperature Room temperature

6) Primary antibody

Concentration or dilution: trials at 1/200, 1/500 and 1/1000 for WB. 1/50, 1/100 and 1/200 for IHC

Diluent buffer WB: Odyssey blocking buffer diluted ½ in PBS 1x. PBS 1x + 0.1% BSA for IHC (also tried 1% BSA and PBST)

Incubation time trials for IHC: 4deg overnight, 2 hours at 37deg and 2 hours at RT.

Incubation temperature:

7) Secondary antibody:

Species: 1/10000 or 1/50000 or 1/2000 Goat anti Rabbit –alexa 633 for IHC and 1/10000 or 1/2000 Donkey anti Rabbit-680 (odyssey) for WB.

Reacts against:

Concentration or dilution

Diluent buffer Diluted in Blocking buffer/PBS (odyssey) for WB or BSA0.1% or 1% in PBS for IHC.

Incubation time 1 hour

Incubation temperature: RT

Fluorochrome or enzyme conjugate: Alexa-633 for IHC (also tried Alexa 488) and 680 (odyssey antibodies)

8) Washing after primary and secondary antibodies: 3 times 5 min with PBS1x for primary and 3 times 5 min PBS 1x for secondary.

Buffer PBS1x

Number of washes 3

9)Detection method Odyssey (WB) and Confocal SP5 Leica (IHC)

10) How many times have you run this staining? 5 times for IHC and 5 times for WB

Do you obtain the same results every time? No, for WB sometimes I get wrong bands and sometimes I don’t get bands

What steps have you altered to try and optimize the use of this antibody? Time and temperature of incubation, dilution of primary and secondary antibody, %BSA

Read More
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.

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. I am not sure why this antibody was showing background/non-specific binding.

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.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up