rev: 5/19/2008
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ANTIBODIES
(anti-Human and others as indicated)
RDI Division of Fitzgerald Industries Intl offers a wide line of antibodies. Since no one antibody works best for all applications (neutralization, blotting, ELISA, etc), we offer many different types of antibodies to help solve this problem. Please inquire for other applications or types of antibodies not listed below.
Cystic Fibrosis Transmembrane Regulator (CFTR)
-see below 4monoclonals and 1 polyclonal
Polyclonal (Rabbit) Anti-Cystic Fibrosis Transmembrane Conductance Regulator Antibody
cat# RDI-CFTRabrX $517.00/100ug
Presentation: 100 ug epitope
affinity purified IgG containing PBS containing 1.0
mg/ml BSA and 0.05% sodium azide. Store at -20 C.
Avoid repeated freeze/thaw cycles.
Immunogen: immunizing peptide corresponds to amino
acid residues 103-117 from human CFTR protein. This sequence is completely
conserved between human, rabbit, and monkey and there is a one amino acid
substitution in rat, bovine, and sheep. The immunizing peptide (cat#RDI-CFTRabrx-CPX $156/50ug) is available for use in
neutralization and control experiments. (Note:Product
cat#RDI-CFTRabrx-CPX, has been discontinued.)
Sequence: G(103) R I I A S Y D P D N K E E R(117)
Use: cytochemsitry: approx 2ug/ml
-detects cystic fibrosis transmembrane conductance factor (CFTR) from cells overexpressing the human protein. It has beeen successfully used in immunocytochemistry procedures. Immunocytochemical staining of HEK293 cells overexpressing human CFTR with this antibody results in staining primarily of the plasma membrane.
References: J. Walker, J. Watson, C. Holmes, A. Edelman and G. Banting. 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 Science, 108: 2433-2444, 1995.
G, Pier, et. al. Salmonella typhi uses CFTR to enter intestinal epithelial cells. Nature: 393: 79-82, 1998.
Backgorund: 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.
For In Vitro research use only
Mouse anti-Human CFTR (N terminus) Clone MM13-4
Predicted Multi-species(Monkey/Rabbit/Sheep/Cow/Mouse)
cat# RDI-CFTRabm-34
Price: $593.25
Package: 200 micrograms of Protein G purified antibody in 0.2ml 0.1M
Tris-glycine,pH7.4, 0.15M NaCl, 0.05% sodium azide before the addition of glycerol to 30%.
Clone: MM13-4
Isotype: mIgG1
Antigen: Synthetic peptide (RKGYRQRLELSD) corresponding to residues 25-36 of
human cystic fibrosis transmembrane conductance
regulator (CFTR). The immunizing sequence is identical in monkey and rabbit,
has 11/12 identical amino acids in sheep, cow and frog, 10/12 identify in mouse
and 9/12 in rat.fusion protein corresponding to
residues 1197-1480 of human cystic fibrosis transmembrane
conductance regulator (CFTR).
Purification: Protein G
Specificity: recognizes human CFTR, Mr 170Kda
Two additional proteins were detected, Mr52 and 38Kda (not ideintified) (other species not determined but predicted
based on antigen homology).
Use: -western blot: approx 0.5ug/ml detects CFTR from CFTR- transfected
BHK and 50-100ug of human T84 colon carcinoma epithelial RIPA cell lysates. CFTR can run as a diffuse protein on SDS-PAGE.
-histochemistry: reported to stain CFTR in T84 cells
-immnoprecipitation: reported to IP CFTR
ref: Kartner, et al, Meth Enzymol 292:629-652, 1998
Storage: Store at -20 Deg C or below. Avoid repeated freeze thaw cycles.
Precautions: For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics.
Immunoblot Protocol
1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a cell lysate sample (cell lysis buffer: 50mM
Tris-HCl, pH 7.4; 1% NP-40; 0.25% sodium deoxycholate; 150mM NaCl; 1mM EGTA; 1mM PMSF; 1ug/ml each aprotinin, leupeptin, pepstatin; 1mM Na3VO4; 1mM NaF) and transfer the proteins to nitrocellulose. Wash the blotted nitrocellulose twice with water.
2. Block the blotted nitrocellulose in freshly prepared PBS containing 3% nonfat dry milk and 0.1% Tween 20 PBST-MLK) for 20 minutes at 20-25°C with constant agitation.
3. Incubate the nitrocellulose with 0.1-0.5ug/ml of anti-CFTR, diluted in freshly prepared PBST-MLK overnight with agitation at 4°C.
4. Wash the nitrocellulose twice with water.
5. Incubate the nitrocellulose in the secondary reagent of choice (a goat anti-mouse HRP conjugated IgG, dilution was used) in PBST-MLK for 1.5 hours at room temperature with agitation.
6. Wash the nitrocellulose with water twice.
7. Wash the nitrocellulose in PBS-0.05% Tween 20 for 3-5 minutes.
8. Rinse the nitrocellulose in 4-5 changes of water.
9. Use detection method of choice (enhanced chemiluminescence was used).
Mouse anti-Human CFTR (C terminus) Clone M3A7
cat# RDI-CFTRabm-A7
Price: $593.25
Package: 200 micrograms of Protein G purified antibody in 0.256ml 0.1M
Tris-glycne,pH7.4, 0.15M NaCl, 0.05% sodium azide before the addition of glycerol to 30%.
Clone: M3A7
Isotype: mIgG1
Antigen: fusion protein corresponding to residues 1197-1480 of human cystic
fibrosis transmembrane conductance regulator (CFTR).
The epitope maps within amino acids 1365-1395.
Purification: Protein G
Specificity: recognizes human CFTR, Mr 170Kda (other
species not determined).
Use: -western blot: approx 0.5ug/ml detects CFTR from CFTR- tranfected
BHK and 50-100ug of human T84 colon carcinoma epithelial RIPA cell lystaes. Do not boil the lysate.
Recommend incubate at 37 DEG C for 30 minutes. CFTR can run as a diffuse
protein on SDS-PAGE.
-histochemsitry: reported to stain CFTR in human pancreatic tissue sections
-immnoprecipitation: reported to IP CFTR
ref: Kartner, et al, Meth Enzymol 292:629-652, 1998
J Biol Chem 274:21873-21877, 1999
Storage: Store at -20 Deg C or below. Avoid repeated freeze thaw cycles.
Precautions: For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics.
Immunoblot Protocol
1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a non- boiled cell lysate sample (cell lysis buffer: 50mM Tris-HCl, pH 7.4; 1% NP-40; 0.25% sodium deoxycholate; 150mM NaCl; 1mM EGTA; 1mM PMSF; 1mg/ml each aprotinin, leupeptin, pepstatin; 1mM Na3VO4; 1mM NaF) and transfer the proteins to nitrocellulose. Wash the blotted nitrocellulose twice with water.
2. Block the blotted nitrocellulose in freshly prepared PBS containing 3% nonfat dry milk and 0.1% Tween 20 (PBST-MLK) for 60 minutes at 20- 25°C with constant agitation.
3. Incubate the nitrocellulose with 0.5ug/ml of anti-CFTR, diluted in freshly prepared PBST-MLK overnight with agitation at 4°C.
4. Wash the nitrocellulose twice with water.
5. Incubate the nitrocellulose in the secondary reagent of choice (a goat anti-mouse HRP conjugated IgG,) in PBST-MLK for 1.0 hour at room temperature with agitation.
6. Wash the nitrocellulose with water twice.
7. Wash the nitrocellulose in PBS-0.05% Tween 20 for 10 minutes.
8. Rinse the nitrocellulose in water for 30 minutes.
9. Use detection method of choice (enhanced chemiluminescence was used).
Mouse anti-Human CFTR (Mid Region) Clone L12B4
cat# RDI-CFTRabm-B4
Price: $562.00/vial $531.00/vial 2 or more
Package: 200 micrograms of Protein G purified antibody in 0.2ml 0.1M
Tris-glycne,pH7.4, 0.15M NaCl, 0.05% sodium azide before the addition of glycerol to 30%.
Clone: L12B4
Isotype: mIgG2a
Antigen: fusion protein corresponding to residues 348-698 of human cystic
fibrosis transmembrane conductance regulator (CFTR).
The epitope maps within amino acids 386-412.
Purification: Protein G
Specificity: recognizes human CFTR, Mr 170Kda (other
species not determined). Also sees two additional protein bands at approx 100
and 140kda.
Use: -western blot: approx 0.5ug/ml detects CFTR from CFTR- transfected
BHK and 50-100ug of human T84 colon carcinoma epithelial RIPA cell lysates. Do not boil the lysate.
Recommend incubate at 37 DEG C for 30 minutes. CFTR can run as a diffuse
protein on SDS-PAGE.
-histochemistry: reported to stain CFTR in human
pancreatic tissue sections
-immnoprecipitation: reported to IP CFTR
ref: Kartner, et al, Meth Enzymol 292:629-652, 1998
J Biol Chem 274:21873-21877, 1999
Storage: Store at -20 Deg C or below. Avoid repeated freeze thaw cycles. Precautions: For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics.
Precautions: For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. We are not responsible for any patent infringements that might result with the use of or derivation of this products.
RDI Division of Fitzgerald Industries Intl
phone (800) 370-2222
or (978) 371-6446 or (800) 370-2222
fax (978) 371-2266
EMAIL:antibodies@fitzgerald-fii.com
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