Overview

  • Product name
  • Description
    Rabbit polyclonal to TLS/FUS
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Dog, Non human primates
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 250 - 350 of Human TLS/FUS.

    Read Abcam's proprietary immunogen policy (Peptide available as ab31610.)

  • Positive control
    • This antibody gave a positive signal in nuclear extracts from HeLa and Jurkat cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab23439 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 Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 73 kDa (predicted molecular weight: 75 kDa).
IP Use at an assay dependent concentration.

Target

  • Function
    Binds both single-stranded and double-stranded DNA and promotes ATP-independent annealing of complementary single-stranded DNAs and D-loop formation in superhelical double-stranded DNA. May play a role in maintenance of genomic integrity.
  • Tissue specificity
    Ubiquitous.
  • Involvement in disease
    Note=A chromosomal aberration involving FUS is found in a patient with malignant myxoid liposarcoma. Translocation t(12;16)(q13;p11) with DDIT3.
    Note=A chromosomal aberration involving FUS is a cause of acute myeloid leukemia (AML). Translocation t(16;21)(p11;q22) with ERG.
    Defects in FUS may be a cause of angiomatoid fibrous histiocytoma (AFH) [MIM:612160]. A distinct variant of malignant fibrous histiocytoma that typically occurs in children and adolescents and is manifest by nodular subcutaneous growth. Characteristic microscopic features include lobulated sheets of histiocyte-like cells intimately associated with areas of hemorrhage and cystic pseudovascular spaces, as well as a striking cuffing of inflammatory cells, mimicking a lymph node metastasis. Note=A chromosomal aberration involving FUS is found in a patient with angiomatoid fibrous histiocytoma. Translocation t(12;16)(q13;p11.2) with ATF1 generates a chimeric FUS/ATF1 protein.
    Defects in FUS are the cause of amyotrophic lateral sclerosis type 6 (ALS6) [MIM:608030]. ALS6 is a familial form of amyotrophic lateral sclerosis. ALS is a neurodegenerative disorder affecting upper motor neurons in the brain and lower motor neurons in the brain stem and spinal cord, resulting in fatal paralysis. Sensory abnormalities are absent. Death usually occurs within 2 to 5 years. The etiology of amyotrophic lateral sclerosis is likely to be multifactorial, involving both genetic and environmental factors. The disease is inherited in 5-10%.
  • Sequence similarities
    Belongs to the RRM TET family.
    Contains 1 RanBP2-type zinc finger.
    Contains 1 RRM (RNA recognition motif) domain.
  • Post-translational
    modifications
    Arg-216 and Arg-218 are dimethylated, probably to asymmetric dimethylarginine.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 75 kDa DNA pairing protein antibody
    • 75 kDa DNA-pairing protein antibody
    • ALS6 antibody
    • Amyotrophic lateral sclerosis 6 antibody
    • fus antibody
    • FUS CHOP antibody
    • Fus like protein antibody
    • FUS_HUMAN antibody
    • FUS1 antibody
    • Fused in sarcoma antibody
    • Fusion (involved in t(12;16) in malignant liposarcoma) antibody
    • Fusion derived from t(12;16) malignant liposarcoma antibody
    • Fusion gene in myxoid liposarcoma antibody
    • Heterogeneous nuclear ribonucleoprotein P2 antibody
    • hnRNP P2 antibody
    • hnRNPP2 antibody
    • Oncogene FUS antibody
    • Oncogene TLS antibody
    • POMp75 antibody
    • RNA binding protein FUS antibody
    • RNA-binding protein FUS antibody
    • TLS antibody
    • TLS CHOP antibody
    • Translocated in liposarcoma antibody
    • Translocated in liposarcoma protein antibody
    see all

Images

  • All lanes : Anti-TLS/FUS antibody (ab23439) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 2 : Jurkat nuclear extract lysate (ab14844)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 75 kDa
    Observed band size: 75 kDa


    Exposure time: 8 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab23439 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

  • Anti-TLS/FUS antibody (ab23439) at 1/1000 dilution + Human 293T cell - whole cell lysate at 30000 cells

    Secondary
    An HRP Goat polyclonal to Rabbit IgG at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 75 kDa
    Observed band size: 72 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 45 kDa (possible non-specific binding), 50 kDa (possible non-specific binding)



    Blocking Step: 5% Milk; 1% BSA for 16 hours at 4°C

    See Abreview

References

This product has been referenced in:
  • Liu C  et al. A cytoplasmic long noncoding RNA LINC00470 as a new AKT activator to mediate glioblastoma cell autophagy. J Hematol Oncol 11:77 (2018). WB, IP . Read more (PubMed: 29866190) »
  • Wang F  et al. The RNA-binding protein QKI5 regulates primary miR-124-1 processing via a distal RNA motif during erythropoiesis. Cell Res 27:416-439 (2017). Read more (PubMed: 28244490) »
See all 17 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - nuclear (hESC)
Gel Running Conditions
Non-reduced Denaturing (4-20%)
Loading amount
20 µg
Specification
hESC
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Mr. Miguel Fidalgo

Verified customer

Submitted Nov 13 2015

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 alternative antibody ab84078:

Order number: #######


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

We have now tried all of your suggestions to optomise the antibody please
see below. We have not had any success at all with any of the options. We
cannot afford to waste any more time or precious time on this and would
like a new antibody. We have probably spent more on reagents than the
original antibody cost in the first place and it is not viable to continue.
Please can you advise where I go from here?
Protocols carried out in the attempt to optimise FUS antibody ab23439 from
abcam:
(nb: Citrate pretreatment: sodium citrate buffer (pH 6.0), microwave at
800W for 5 minutes with lid on, cool for 5 minutes with lid off, microwave
at 800W for 5 minutes, cool gently, wash.
EDTA pretreatment: 3 minutes in formic acid, microwave at 800W for 10
minutes in 1mM EDTA, cool gently, wash)

1. Pretreatment: EDTA, methanol block
Antibody concentration: 1:300, 1:400
Optimisation procedure: Overnight incubation at room temp.

2. Pretreatment: Citrate, methanol block
Antibody concentration: 1:100, 1:200, 1:300, 1:400, 1:500, 1:1000, 1:2000
Optimisation procedure: Overnight incubation at room temp.

3. Pretreatment: Citrate, methanol block
Antibody concentration: 1:1000
Optimisation procedure: Permeabilisation in 0.2% Triton in PBS 10 mins.
Overnight incubation at room temp.

4. Pretreatment Citrate, methanol block
Antibody concentration 1:1000
Optimisation procedure: 0.2% Tween in antibody dilution buffer and wash
buffer. Overnight incubation at room temp.

5. Pretreatment: Citrate, methanol block
Antibody concentration: 1:1000
Optimisation procedure: Permeabilisation in 0.2% Triton in PBS 10 mins.
0.2% Tween in antibody dilution buffer and wash buffer. Overnight at room
temp.

6. Pretreatment: Citrate, methanol block
Antibody concentration: 1:4000
Optimisation procedure: Overnight incubation at 4°C

7. Pretreatment: Citrate, methanol block
Antibody concentration: 1:4000
Optimisation procedure: Permeabilisation in 0.2% Triton in PBS 10 mins.
0.2% Tween in antibody dilution buffer and wash buffer. Overnight
incubation at 4°C.

8. Pretreatment: Citrate
Antibody concentration: 1:1000
Optimisation procedure: No methanol block, straight to wash. Overnight
incubation at room temp.

9. Pretreatment: Citrate
Antibody concentration: 1:1000
Optimisation procedure: No methanol block, straight to wash.
Permeabilisation in 0.2% Triton in PBS 10 mins. Overnight incubation at
room temp.

10. Pretreatment: Citrate
Antibody concentration: 1:1000
Optimisation procedure: No methanol block, straight to wash. 0.2% Tween in
antibody dilution buffer and wash buffer. Permeabilisation in 0.2% Triton
in PBS 10 mins. Overnight incubation at room temp.

11. Pretreatment: Citrate,
Antibody concentration: 1:4000
Optimisation procedure: No methanol block, straight to wash.
Permeabilisation in 0.2% Triton in PBS 10 mins. 0.2% Tween in antibody
dilution buffer and wash buffer. Overnight incubation at 4°C.

12. Pretreatment: Citrate,
Antibody concentration: 1:4000
Optimisation procedure: No methanol block, straight to wash. Overnight
incubation at 4°C.

Best wishes,

Read More
Answer

Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments, and would like to offer a free of charge replacement or credit note in compensation (providing the product has been purchased in the last 6 months). In order to arrange this, I would appreciate if you could confirm your order number and date of purchase? Was this Abcam order ref 932379 (PO FMDY1090259) ordered by Joanna McTeirman on 12th Aug?

Thank you for your continued cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (293T Cells)
Loading amount
30000 cells
Specification
293T Cells
Gel Running Conditions
Reduced Denaturing (4-20%)
Blocking step
5% Milk, 1% BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Feb 12 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HEK293 overexpressing HA-tagged FUS)
Total protein in input
2e+006 cells
Specification
HEK293 overexpressing HA-tagged FUS
Immuno-precipitation step
Other - Dynabeads

Abcam user community

Verified customer

Submitted Oct 14 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Loading amount
30 µg
Specification
HEK293
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 07 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblast)
Loading amount
50 µg
Specification
Mouse embryonic fibroblast
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 15 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up