rev:       january 11, 2004

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  ANTIBODIES  

(anti-Human and others as indicated)

RDI Divison of researchd 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    $469.00/100ug

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: $562.00/vial   $531.00/vial 2 or more

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: $562.00/vial   $531.00/vial 2 or more

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 Divison of researchd Industries Intl

San Jose, 95123 CA Snell ave 658

USA

phone (800) 370-2222

      or (978) 371-6446 or (800) 370-2222

EMAIL:margaret@cellular-research.com

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