Overview

  • Product name
    Anti-CFTR antibody [CF3]
    See all CFTR primary antibodies
  • Description
    Mouse monoclonal [CF3] to CFTR
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC/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, 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

  • Form
    Liquid
  • Storage instructions
    Shipped 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 buffer
    Preservative: 0.05% Sodium azide

    Diluted ascites
  • Concentration information loading...
  • Purity
    Ascites
  • 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 number
    CF3
  • Isotype
    IgM
  • 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 1/200.
WB 1/500. Predicted molecular weight: 168 kDa.
Inhibition Assay Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab91545 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Involved in the transport of chloride ions. May regulate bicarbonate secretion and salvage in epithelial cells by regulating the SLC4A7 transporter.
  • Tissue specificity
    Found on the surface of the epithelial cells that line the lungs and other organs.
  • Involvement in disease
    Defects 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 similarities
    Belongs 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.
  • Domain
    The 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 localization
    Early 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 (ATP-binding cassette sub family C, member 7) 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

Images

  • 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.

  • 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.
  • 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.

  • 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.

  • Immunocytochemistry/Immunofluorescent analysis of HeLa cells labelling CFTR with ab2784. 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.

  • 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.

  • 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.

  • 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.

References

This product has been referenced in:
  • Tomati V  et al. Thymosin a-1 does not correct F508del-CFTR in cystic fibrosis airway epithelia. JCI Insight 3:N/A (2018). Read more (PubMed: 29415893) »
  • Carbone A  et al. Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells. Stem Cells Int 2018:1203717 (2018). ICC/IF . Read more (PubMed: 29531530) »
See all 27 Publications for this product

Customer reviews and Q&As

1-10 of 18 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate buffer pH6
Permeabilization
No
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Nov 28 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (muscle)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate buffer pH6
Permeabilization
No
Specification
muscle
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Nov 28 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin carcinoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
skin carcinoma
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Nov 21 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Nov 21 2017

Abreviews
Application
CyTOF
Sample
Mouse Cell (Lung)
Specification
Lung

Irit Milman

Verified customer

Submitted Nov 14 2017

Answer

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 permiabilization is used. I think this may be due to the epitope recognised by the antibody not being accessible unless this is used. Although the immunogen used is from the region 103-117, which does correspond to an extracellular loop according to Uniprot (reference P13569) this does not necessarily mean this is accessible to the antibody. It may be that the detergent only allows the antibody sufficient access to this loop. It may be worth trying the experiment with a different cell line if you have access to this to see if the same is observed.

I can understand that the point in your experiment was to be able to detect only the membrane bound protein and thus this makes the antibody incompatible with these aims. I cannot find another antibody in our catalogue that would be likely to be able to work without permiabilisation. I am sorry that I could not be of more help in this instance.

If you have any further questions, please do not hesitate to ask.

Read More

Answer



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 Ihnen, dass Sie uns darauf aufmerksam gemacht haben.

Laut dem Labor wird diese Antikörperkonzentrationüber eineA280 nm Absorptionkurve gemessen.

Read More

Question
Answer

Thank you for contacting us.



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 sorry we do not sell the peptide as a standalone product.





I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.


Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

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 extracellular domain of the antigen and thus this antibody is detecting an extracellular domain of the CFTR protein.

Unfortunately, they don’t have any data of this antibody being used on live cells. We cannot confirm that this antibody works under these conditions, as we have not validated it in-house nor do we have a publication that stated they used it successfully in this type of application.

You could try using the antibody on fixed cells to see if you would get any staining in those circumstances, as that wil at least let you know if the antibody is recognizing the correct target or not.

Please let me know if there is anything else I can help you with.

Read More

Answer

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 information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

We wish you the best of luck with your research and look forward to a reply.

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1-10 of 18 Abreviews or Q&A

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