Overview

  • Product nameAnti-CFTR antibody [CF3]See all CFTR primary antibodies ...
  • Description
    Mouse monoclonal [CF3] to CFTR
  • SpecificityWe have received feedback from users that suggests this antibody might not work with unfixed, unpermeabilized cells. We only test with fixed cells and can only guarantee ab2784 to work with fixed cells.
  • Tested applicationsICC/IF, IP, IHC-P, WB, Inhibition Assay, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Guinea pig, Human, Pig
    Predicted to work with: Sheep, Rabbit, Horse, Cow, Dog, Chimpanzee, Non Human Primates, Rhesus monkey, Gorilla, Loxodonta Africana (African bush elephant)
  • Immunogen

    Synthetic peptide corresponding to Human CFTR aa 103-117. Found in the first extracellular loop of human and rabbit CFTR.
    Sequence:

    GRIIASYDPDNKEER


    (Peptide available as ab4911)

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferPreservative: 0.05% Sodium azide

    Diluted ascites
  • Concentration information loading...
  • PurityAscites
  • Primary antibody notes Cystic Fibrosis (CF) is a common lethal genetic disease caused by mutations of the gene coding for the cystic fibrosis transmembrane conductance factor, a cAMP regulated chloride channel. Approximately 70% of all CF cases share the deletion of a phenylalanine at position 508 (delta F508) which results in abnormal chloride transport. Since the CF mutation is lethal, most often by lung and liver disease, it raises the question of why this genetic disease remains as common as it is. One possible explanation is that Salmonella typhi has been shown to use CFTR to enter intestinal epithelial cells and that delta F508 heterozygote and homozygote mice showed 86% and 100% reductions in S.typhi intestinal submucosal uptake.
  • Clonality Monoclonal
  • Clone numberCF3
  • IsotypeIgM
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab2784 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 at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 168 kDa.
Inhibition Assay Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

Target

  • FunctionInvolved in the transport of chloride ions. May regulate bicarbonate secretion and salvage in epithelial cells by regulating the SLC4A7 transporter.
  • Tissue specificityFound on the surface of the epithelial cells that line the lungs and other organs.
  • Involvement in diseaseDefects in CFTR are the cause of cystic fibrosis (CF) [MIM:219700]; also known as mucoviscidosis. CF is the most common genetic disease in the Caucasian population, with a prevalence of about 1 in 2'000 live births. Inheritance is autosomal recessive. CF is a common generalized disorder of exocrine gland function which impairs clearance of secretions in a variety of organs. It is characterized by the triad of chronic bronchopulmonary disease (with recurrent respiratory infections), pancreatic insufficiency (which leads to malabsorption and growth retardation) and elevated sweat electrolytes.
    Defects in CFTR are the cause of congenital bilateral absence of the vas deferens (CBAVD) [MIM:277180]. CBAVD is an important cause of sterility in men and could represent an incomplete form of cystic fibrosis, as the majority of men suffering from cystic fibrosis lack the vas deferens.
  • Sequence similaritiesBelongs to the ABC transporter superfamily. ABCC family. CFTR transporter (TC 3.A.1.202) subfamily.
    Contains 2 ABC transmembrane type-1 domains.
    Contains 2 ABC transporter domains.
  • DomainThe PDZ-binding motif mediates interactions with GOPC and with the SLC4A7, SLC9A3R1/EBP50 complex.
  • Post-translational
    modifications
    Phosphorylated; activates the channel. It is not clear whether PKC phosphorylation itself activates the channel or permits activation by phosphorylation at PKA sites.
    Ubiquitinated, leading to its degradation in the lysosome. Deubiquitination by USP10 in early endosomes, enhances its endocytic recycling.
  • Cellular localizationEarly endosome membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ABC 35 antibody
    • ABC35 antibody
    • ABCC 7 antibody
    • ABCC7 antibody
    • ATP binding cassette sub family C member 7 antibody
    • ATP Binding Cassette Superfamily C Member 7 antibody
    • ATP binding cassette transporter sub family C member 7 antibody
    • ATP-binding cassette sub-family C member 7 antibody
    • cAMP dependent chloride channel antibody
    • cAMP-dependent chloride channel antibody
    • CF antibody
    • CFTR antibody
    • CFTR/MRP antibody
    • CFTR_HUMAN antibody
    • Channel conductance controlling ATPase antibody
    • Channel conductance-controlling ATPase antibody
    • Cystic fibrosis transmembrane conductance regulator antibody
    • Cystic fibrosis transmembrane conductance regulator ATP binding cassette sub family C member 7 antibody
    • Cystic Fibrosis Transmembrane Regulator antibody
    • dJ760C5.1 antibody
    • MRP 7 antibody
    • MRP7 antibody
    • TNR CFTR antibody
    see all

Anti-CFTR antibody [CF3] images

  • ab2784 staining CFTR in human pancreatic cells by Immunocytochemistry/ Immunofluorescence. The cells were paraformaldehyde fixed, permeabilised in 0.5% Triton X-100. Samples were then incubated with primary antibody at 1/50 for 24 hours at 4°C. The secondary antibody used was an anti-mouse IgG conjugated to FITC (green) used at a 1/200 dilution.
  • Ab2784 staining Human normal colon. Staining is localized to the cell membrane.
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • Overlay histogram showing A549 cells stained with ab2784 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2784, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in A549 cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Immunofluorescent analysis of CFTR using CFTR Monoclonal antibody (CF3) ab2784 shows staining in WiDr colon carcinoma cells. CFTR staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing CFTR ab2784 at a dilution of 1:100-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunofluorescent analysis of CFTR using CFTR Monoclonal antibody (CF3) ab2784 shows staining in U251 glioma cells. CFTR staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing CFTR ab2784 at a dilution of 1:100-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunofluorescent analysis of CFTR using CFTR Monoclonal antibody (CF3) ab2784 shows staining in HeLa cells. CFTR staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing CFTR ab2784 at a dilution of 1:100-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunohistochemistry was performed on cancer biopsies of deparaffinized Human colon carcinoma tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing CFTR ab2784 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Human tonsil tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing CFTR ab2784 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Human pancreas tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing CFTR ab2784 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

References for Anti-CFTR antibody [CF3] (ab2784)

This product has been referenced in:
  • Villella VR  et al. Disease-relevant proteostasis regulation of cystic fibrosis transmembrane conductance regulator. Cell Death Differ 20:1101-15 (2013). Read more (PubMed: 23686137) »
  • Monterisi S  et al. CFTR regulation in human airway epithelial cells requires integrity of the actin cytoskeleton and compartmentalized cAMP and PKA activity. J Cell Sci 125:1106-17 (2012). Human . Read more (PubMed: 22302988) »

See all 11 Publications for this product

Product Wall

Thank you for contacting us and reporting the problems you have encountered in using the Anti-CFTR antibody [CF3] (ab2784).

I have tried to seek further information in order to understand why the antibody only appears to be working when permia...

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Uns ist tatsächlich ein Datenblattfehler unterlaufen: Da es sich hier um einen Ascitis- Antikörper handelt, ist dieser natürlich nicht aufgereinigt. Wir werden das Datenblatt dementsprechend ändern, und bedanken uns bei Ih...

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The Anti-CFTR antibody [CF3] immunizing peptide corresponds to amino acid residues 103-117 (G(103) R I I A S Y D P D N K E E R(117) found in the first extracellular loop of human and rabbit CFTR. I am...

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Thank you for contacting Abcam.

I have talked to the lab and they have confirmed that the immunogen for this antibody is amino acids ********of CFTR, Cystic Fibrosis Transmembrane Conductance Regulator. Therefore, the immunogen was from an ex...

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Thank you for your previous email letting us know about the trouble with ab2784. Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested informatio...

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Thank you for contacting Abcam. Under our Abpromsie, ab2784 is already guaranteed to work in WB and IF, in both human and mouse tissue. Therefore it is not eligible for a testing discount. Please let me know if there is anything else I can help you with.

Thank you for contacting us and letting us know about the trouble with ab2784. I do have a couple of additional questions, so that I better understand the situation: 1) Does the polymer detection system work with mouse IgM antibodies? Many secondary an...

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Thank you for your call yesterday and for your patience while I have been in touch with the lab regarding ab2784. The Western blot application was added to the datasheet based on characterization by the scientists who originally created clone CF...

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Application Flow Cytometry
Sample Human Cell (tracheobronchial epithelial-CFTE(F508d)-HTE (CF))
Specification tracheobronchial epithelial-CFTE(F508d)-HTE (CF)
Preparation Cell harvesting/tissue preparation method: Cells scraped from 24 well plates and washed in PBS (Trypsin was used initially to remove cells but change for final experiments to scraping them off).
Sample buffer: 0.1% BSA in PBS
Fixation Paraformaldehyde
Permeabilization No
Gating Strategy Only one cell population which was gated using Expo Analysis software
Username

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Verified customer

Submitted Feb 02 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Pancreatic tissue primary culture)
Specification Pancreatic tissue primary culture
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton x100
Blocking step (agent) for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Oct 13 2008

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