Overview

  • Product name
  • Description
    Rabbit polyclonal to CFTR
  • Host species
    Rabbit
  • Specificity
    Detects cystic fibrosis transmembrane conductance factor (CFTR) from cells overexpressing the human protein.
  • Tested applications
    Suitable for: ICC/IF, IP, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Rat, Sheep, Rabbit, Guinea pig, Cow, Dog, Pig, Chimpanzee, Monkey, Non human primates, Cynomolgus monkey, Rhesus monkey, Gorilla, Elephant
  • Immunogen

    Synthetic peptide corresponding to Human CFTR aa 103-117.
    Sequence:

    GRIIASYDPDNKEER

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
    Constituents: 99% PBS, 0.1% BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • 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
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab2916 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 1 µg/ml.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 168 kDa.

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

  • Immunolocalization of CFTR in HEK293 cells stably transfected with the CFTR gene using ab2916.

References

This product has been referenced in:
  • Yang X  et al. High CFTR expression in Philadelphia chromosome-positive acute leukemia protects and maintains continuous activation of BCR-ABL and related signaling pathways in combination with PP2A. Oncotarget 8:24437-24448 (2017). WB . Read more (PubMed: 28445932) »
  • Klein T  et al. Investigation and characterization of the duct cell-enriching process during serum-free suspension and monolayer culture using the human exocrine pancreas fraction. Pancreas 38:36-48 (2009). Read more (PubMed: 18665014) »
See all 2 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Question

A customer just purchased #ab2916 and did immunofluorescence. Unfortunately, he cannot get staining fluorescence. Below is the questionnaire. Thank you. 1. Order details: * Batch number: ab2916 * Antibody storage conditions (temperature/reconstitution etc) -20? 2. Please describe the problem (high background, no staining etc). no staining 3. On what material are you testing the antibody in IHC * Species: human * Cell Iine or tissue: sperm 4. How did you fix the samples? * Ethanol, methanol: (was it ice cold) * Acetone: (was it ice cold) * Paraformaldehyde (percentage, perfusion fixed on not): 4%, not perfusion fixed * Fixation time: 40min * Fixation temperature: room temperature 6. Blocking steps: * For HRP detection method: did you block endogenous peroxidases: * How did you block the unspecific binding sites: BSA * For how long: 1h 7. Primary antibody * Specification (in which species was it raised against): rabbit * At what dilution(s) have you tested this antibody: 1:500 * What dilution buffer was used: BSA * Incubation time: 16h * Incubation temperature: 4? * What washing steps were done: PBS 5min, 4times 8. Secondary antibody * Specification (in which species was it raised against): goat * What dilution buffer was used: BSA * Incubation time: 1h * Incubation temperature: room temperature * What washing steps were done: PBS 5min, 4times * Do you know whether the problems you are experiencing come from the secondary? No, not the problem from 2nd antibody 9. What detection method are you using? immunofluorescence 10. Background staining See attachment 11. Which detection system did you use? FITC 12. Did you apply positive and negative controls along with the samples? We applied negative controls, and it’s reported CFTR expressed by sperm and could be detected by immunofluorescence. 13. Optimization attempts * How many times have you tried the IHC? One time * Do you obtain the same results every time? yes * What steps have you altered?

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Answer

I'm very sorry to hear one of your customers is experiencing problems with ab2916, indeed CFTR is known to be present in sperm and should be detectable with ab2916. I think the problem comes from the fixation which is damaging the epitopes. I would recommend to fix the cells in methanol for 5 minutes at -20C. I enclose below the protocol used by Walker et al (Walker J et al. Production and characterisation of monoclonal and polyclonal antibodies to different regions of the cystic fibrosis transmembrane conductance regulator (CFTR): detection of immunologically related proteins. J Cell Sci 108 (Pt 6):2433-44 (1995)): Wash cells 3x PBS, then fixe and permeabilize by incubation in methanol at -20°C for 5 min. Incubate coverslips in PBS/0.2% BSA for 5 min followed by a PBS wash and incubation with primary antibody in PBS/0.2% BSA for 1 h at room temperature. After washing in PBS/0.2 % BSA to remove excess primary antibody, incubate cells in secondary antibody. Please let me know if the customer still has problems with this protocol and I can offer you a replacement vial or credit note,

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Answer

Thank you for contacting us for some advice on ab2916 and ab2784. You are correct, both antibodies were made using the same immunizing peptide, however they are raised in different species and more importantly we do not know if they recognise the same epitope within aa103-117, so a different epitope could explain the difference. We have not tested ab2916 in WB and from your feedback it does appear that it does not work in that application, would you please consider leaving a review on this antibody so we can add this information for future users? We reward reviews (negative and positive) with 100 Abcam points which can be used for a number of gifts. I hope the above information will help you, please do not hesitate to contact me again if you require further assistance,

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Answer

Both ab2916 and ab2784 are made from the same synthetic peptide corresponding to amino acid residues 103-117. Ab2784 corresponds to amino acid residues 103-117 found in the first extracellular loop of human and rabbit CFTR. So, it appears that ab2916 was made from the extracellular portion. I hope this information helps; if you have any additional questions, please contact us again.

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Answer

I'm sorry to hear you are experiencing problems with ab2916. This antibody has not been tested for western blotting, it is therefore difficult to find out what the problem might be. A litterature search about CFTR enabled me to find that the lysis buffer used to extract the protein from K562 cells is the following NP40 based buffer: 50 mM HEPES, 250 mM NaCl, 1 mM EDTA, 1% NP-40, 1 mM PMSF, and 1 mM DTT. This information was from the reference "CFTR in K562 human leukemic cells. Assef YA, Damiano AE, Zotta E, Ibarra C, Kotsias BA. Am J Physiol Cell Physiol. 2003 Aug;285(2):C480-8." The article continues: "Nuclei and unbroken cells were removed with centrifugation (6,000 g for 15 min at 4°C). Total protein in each sample was quantified using the bicinchoninic acid assay (BCA Protein, Pierce Chemical). For Western blot studies, 100 ug of total proteins from K562 cells and 50 ug of total proteins from T84 cells were dissolved in loading buffer (4% sodium dodecil sulfate, 0.125 M Tris-HCl, pH 6.8, 0.2 M dithiothreitol, 0.02% bromophenol blue, and 20% glycerol). The preparation was heated to 90°C for 2 min, resolved on 7.5% polyacrylamide gel, and electrotransferred onto nitrocellulose membranes(Hybond ECL, Amersham Pharmacia Biotech, UK)." I would recommend trying a range of dilutions (e.g. 1/100-1/1000) and incubating the antibody overnight at 4C. I hope the above information will help you, please let me know if you require further assistance,

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