Overview

  • Product name
  • Description
    Rabbit polyclonal to PTRF
  • Tested applications
    Suitable for: IP, IHC-P, ICC/IF, ELISA, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, African green monkey, Chinese hamster
    Predicted to work with: Dog
  • Immunogen

    A region within synthetic peptide: PEKEGDELGE GERPEDDTAA IELSSDEAVE VEEVIEESRA ERIKRSGLRR, corresponding to internal sequence amino acids 181-230 of Mouse PTRF

  • Positive control
    • NIH3T3 lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab48824 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
IP 1/200 - 1/250. (for 20 hrs - total 250 ug protein)
IHC-P Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/100.
ELISA Use at an assay dependent concentration.
WB Use a concentration of 0.625 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 43 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.

Target

  • Function
    Plays an important role in caveolae formation and organization. Required for the sequestration of mobile caveolin into immobile caveolae. Termination of transcription by RNA polymerase I involves pausing of transcription by TTF1, and the dissociation of the transcription complex, releasing pre-rRNA and RNA polymerase I from the template. PTRF is required for dissociation of the ternary transcription complex.
  • Involvement in disease
    Congenital generalized lipodystrophy 4
  • Sequence similarities
    Belongs to the PTRF/SDPR family.
  • Post-translational
    modifications
    Phosphorylated. Present in active and inactive forms. Changes in phosphorylation pattern may alter activity.
    Five truncated forms are found in the caveolae. These are thought to be the result of proteolysis and may be phosphorylation-dependent.
  • Cellular localization
    Membrane > caveola. Cell membrane. Microsome. Endoplasmic reticulum. Cytoplasm > cytosol. Mitochondrion. Nucleus. Found at the surface of the caveolae. Also found in the plasma membrane, microsomal and cytosolic fractions and at a low level in the mitochondrial and nuclear fractions. Translocates to the cytoplasm from the caveolae upon insulin stimulation. CAV1 is necessary to recruit it to the cell membrane (By similarity). Co-localizes with CAV1 in lipid rafts in adipocytes.
  • Information by UniProt
  • Database links
  • Alternative names
    • 2310075E07Rik antibody
    • AW546441 antibody
    • Cavin 1 antibody
    • Cavin antibody
    • Cavin-1 antibody
    • CAVIN1 antibody
    • CGL4 antibody
    • FKSG13 antibody
    • FLJ90031 antibody
    • MGC118550 antibody
    • OTTMUSP00000002049 antibody
    • Polymerase I and transcript release factor antibody
    • Ptrf antibody
    • PTRF_HUMAN antibody
    • RNA polymerase I and transcript release factor antibody
    • RP23-279L23.6 antibody
    • TTF I interacting peptide 12 antibody
    see all

Images

  • Anti-PTRF antibody (ab48824) at 1/1000 dilution + HeLa cell lysate at 10 µg

    Secondary
    HRP-conjugated goat anti-rabbit IgG polyclonal at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 43 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 seconds

    This image is courtesy of an anonymous Abreview

    Blocked with 5% milk for 1 hour at 22°C.

    Incubated with the primary antibody for 1 hour at 22°C.

    See Abreview

  • ab48824 (4µg/ml) staining PTRF in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear and cytoplasmic staining.
    Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Immunocytochemical analysis of MDCK (Madin-Darby Canine Kidney) epithelial cells, labeling PTRF with ab48824. Cells were paraformaldehyde fixed, permeabilized in 0.1% Trition in PBS and blocked in 10% serum for 30 mintues at 22°C. Incubation with anti-PTRF diluted 1/300 was for 1 hour at 22°C. Nuclei have been stained blue.

    See Abreview

  • Anti-PTRF antibody (ab48824) at 1/2000 dilution + Rat C6 cells, whole cell lysate at 25 µg

    Secondary
    HRP conjugated goat anti-rabbit antibody at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 43 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

  • Immunocytochemical analysis of HeLa cells, labeling PTRF with ab48824. Cells were paraformaldehyde fixed, permeabilized in 0.1% Trition in PBS and blocked in 10% serum for 30 mintues at 22°C. Incubation with anti-PTRF diluted 1/300 was for 1 hour at 22°C. Nuclei have been stained blue.

    See Abreview

  • All lanes : Anti-PTRF antibody (ab48824) at 1/1000 dilution

    Lane 1 : Mouse embryonic fibroblast lysate
    Lane 2 : Mouse PTRF-KO embryonic fibroblast lysate
    Lane 3 : Mouse PTRF-KO embryonic fibroblast lysate + FLAG-PTRF protein

    Lysates/proteins at 50 µg per lane.

    Secondary
    HRP conjugated Donkey anti-Rabbit at 1/10000 dilution

    Predicted band size : 43 kDa
  • Immunocytochemical analysis of human AT2 cells, labeling PTRF with ab48824. Cells were paraformaldehyde fixed, blocked in 3% BSA and 0.1% saponin in PBS, then incubated with ab48824.

  • Immunohistochemical analysis of FFPE mouse vessel and adipocytes (skeletal muscle) tissue, labeling PTRF with ab48824 (5µg/ml).

References

This product has been referenced in:
  • Zhu B  et al. Similar regulatory mechanisms of caveolins and cavins by myocardin family coactivators in arterial and bladder smooth muscle. PLoS One 12:e0176759 (2017). WB . Read more (PubMed: 28542204) »
  • Ludwig A  et al. Caveolae provide a specialized membrane environment for respiratory syncytial virus assembly. J Cell Sci 130:1037-1050 (2017). Read more (PubMed: 28154158) »

See all 25 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (Pericytes)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Specification
Pericytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Verified customer

Submitted Feb 19 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Dog Cell (MDCK)
Permeabilization
Yes - 0.1% Triton in PBS
Specification
MDCK
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 27 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (pericytes)
Permeabilization
Yes - 0.1% Triton in PBS
Specification
pericytes
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 18 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (cos-7)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
20 µg
Specification
cos-7
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

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Verified customer

Submitted Dec 07 2015

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Brain endothelial cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Brain endothelial cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

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Verified customer

Submitted Jul 29 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (Hela cells)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
Hela cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

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Verified customer

Submitted Jul 29 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (bEnd.3)
Permeabilization
Yes - 0.1% Triton in PBS
Specification
bEnd.3
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 28 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hela)
Permeabilization
Yes - 0.1% Triton in PBS
Specification
Hela
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 28 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Rat Cell lysate - whole cell (L6 cell line)
Total protein in input
200 µg
Specification
L6 cell line
Immuno-precipitation step
Protein A/G
Username

Abcam user community

Verified customer

Submitted Jun 04 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Chinese Hamster Cell lysate - whole cell (CHO cell line)
Total protein in input
200 µg
Specification
CHO cell line
Immuno-precipitation step
Protein A/G
Username

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Verified customer

Submitted Jun 04 2009

1-10 of 24 Abreviews or Q&A

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