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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, histochemistry, 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.


Ankyrin & Ankyrin G abs

Mouse anti-ANKYRIN Monoclonal Antibody


Catalog#: RDI-ankyrabmX    Price:$531.00/50UG VIAL


Background: -Ankyrin consists of a single 215Kda peptide known to be involved in the attachment of spectrin (or fodrin) containing membrane skeleton to the plasma membranes. Ankyrin belongs to a family of closely related polypeptides associated with the plasma membrane of cells in a variety of cell types (e.g. lymphocytes, platelets, fibroblasts and endothelial tissues). Ankyrin has been shown to underlie membrane proteins including CD44, the voltage-dependent sodium channel, NA+/K+ ATPase and the anion exchanger protein. -Functional diversity between members of the ankyrin family is generated by the expression of multiple genes as well as alternative splicing genes as well as alternative splicing of pre-mRna's. It is believed that the formation of a direct connection between ankyrin and functionally important transmembrane proteins/membrane skeleton may be one of the earliest events to occur during signal transduction and cell activation.


Package Size: 100ul IgG2a (0.5mg/ml) in 10mM SODIUM PHOSPHATE, 150mM NaCl, 0.01% NaN3, pH7.5

Isotype: IgG2a (purified from culture supernatant of ankyrin hybridoma cell line)

Antigen: purified human erythroid cell ankyrin

Specificity: reacts specifically with human intracellular ankyrin (215 kDa) in human, mouse, rabbit, rat and bovine cells and tissues. (for histochemistry, requires permeabilization steps as 0.1% saponin)

Use: -western blot (approx 1:1000) with 2-3 hr RT or overnight 4 DEg C with chemiluminescence detection

         -immunoprecipitation

         -cytochemistry, 2% PFA fixation

        -flow cytometry (with permabilization)

       -ELISA


ref: -Bennet, V. Physiological Reviewers, 70:1029- 1054

     -Srinivasan, Y. et al, Nature, 333:177-180, 1988.

     -Morrow, J.S. et al, J. Cell Bio. 108:455-465, 1989

     -Bourguignon, L.Y.W. et al, J. Biol. Chem, 266:11761-11765, 1991

     -Lokeshwar, V.B. et al, Int. Rev. Cytol., 87, 17983-17989, 1991

    -Bourguignon, L.Y.W. et al, Int. Rev Cytol, 87:195-224, 1984.



Storage: Store at -20 DEG C or lower. AVOID FREQUENT   FREEZE THAW CYCLES. Do not freeze diluted   antibody.

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 product.


Mouse anti-Ankyrin G

cat# RDI-ANKYRGabmX    $438.00/100ug

Presentation: 100ug in 1ml(1mg/ml) PBS + 0.2% gelatin and 0.1% sodium azide.

Immunogen: a synthetic peptide derived from the human Ankyrin G spectrin binding domain

Clone: 463

Isotype: mIgG

Reactivity: By Western blot, recognizes the 190kDa isoform of Ankyrin G. This antibody has been shown to work in immunofluorescence applications on frozen tissue. AnlyrinG is a good immunohistochemical marker for axon inital segments, nodes of Ranvier, and epithelial cell-cell contract sites.

Species   Reactivity: human, mouse, rat and dog

Uses: -Western blot: 1-3ug/ml (2-3 hours RT or overnight 4 DEG C with chemiluminescence detection) pos control mouse or rat brain extract

-immunofluorescence (frozen tissue) 2-5ug/ml (pos control unmyelinated axons) using FITC detection

-histochemistry (frozen sections)

Suggestion Trial Protocol (must optimize for each method)

Note: The antibody is sensitive to fixation conditions. The following conditions are recommended:1%PFA for perfusion and 4 hours post-fixation (after perfusion). For sciatic nerve, 2% paraformaldehyde is recommended for 2 hours at 4 DEG C.

-for immunofluorescence:

a) permeabilize for 15min at 4 DEG C in PBS containing 0.5% Triton X-100, 0.2% Tween 20 and 10mg/ml BSA.

b) Wash with PBS + 0.2% Tween-20 3 times at 4DEGC

C)Apply antibody in blocking buffer (PBS with 0.2% Tween-20 and 10mg/ml BSA)

d)Wash with wash buffer (PBS with 0.2% Tween-20) 3-5 times at 4 DEG C

e) Apply secondary antibody (recommend using a biotin labelled secondary) dissolved in PBS with 0.2% Tween-20

f) Wash with wash buffer (PBS with 0.2% Tween-20) 3-5 times at 4 DEG C

g) Apply Streptavidin:FITC for 2 hours at 4 DEG C

h) Wash with wash buffer (PBS with 0.2% Tween-20) 3-5 times at 4 DEG C.

i) Mount slides.

ref: -J. Biol Chem. 270(5)2352-2359 (1995)

       -J. Cell Sci 111:2197-2207 (1998)

Storage: 2-8 deg c (up to one month) or aliquot and store at -20 DEG C in non-frost free freezers. Avoid frequent freeze thaw cycles.

Background:

Ankyrins are a family of spectrin-binding proteins that couple the spectrin/actin cytoskeletal network to the cytoplasmic domains of integral membrane proteins which include channels such as the anion exchanger, voltage-dependent sodium channel, and the Na/K ATPase, as well as neuronal cell adhesion molecules related to L1 . In the nervous system, three different ankyrins are known to exist; ankyrinB, ankyrinG,and ankyrinR. All three ankyrins contain an N-terminal membrane binding domain, an internal domain capable of binding spectrin, and a C-terminal regulatory domain which is the target of alternative splicing. AnkyrinB,the predominant neuronal ankyrin includes two isoforms of 220 kDa and 440 kDa . The 220 kDa alternatively spliced isoform is the major form present in the adult brain, while the 440 kDa form is preferentially expressed in the developing neonatal brain . Three forms of ankyrinG (190 kDa, 270 kDa, 480 kDa) are localized in axonal initial segments and the nodes of Ranvier of myelinated axons where fluxes of action potential occur and ion channels are enriched . Ankyrins comprise approximately 0.5%-1% of the total membrane protein in the adult vertebrate brain .

For Research Use Only


Mouse anti-Ankyrin B

cat# RDI-ANKYRBabmX   $438.00/100ug

Presentation: 100ug in 1ml(1mg/ml) PBS + 0.2% gelatin and 0.1% sodium azide.

Immunogen: a synthetic peptide derived from the human Ankyrin B spectrin binding domain

Clone: 2.20

Isotype: mIgG1

Reactivity: By Western blot, recognizes all spliced forms of ankyrinB which contain a spectrin binding domain. In western blots using mouse or rat brain extract from adult, this antibody detects three isoforms at molecular weights of 440kDa, 2000kDa, and 150kDa. This antioidy has been shown to work in immunofluorescence applications on frozen tissue.

Species   Reactivity: human, mouse, rat

Uses: -Western blot: 1-3ug/ml (2-3 hours RT or overnigth 4 DEG C with chemiluminescence detection) pos control mouse or rat brain extract

          -immunofluorescence (frozen tissue) 2-5ug/ml (pos control unmyelinated axons) using FITC detection

         -histochemistry (frozen sections)


RDI Division of Fitzgerald Industries Intl

34 Junction Square Drive

Concord MA 01742-3049

USA

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

fax     (978) 371-2266

EMAIL:antibodies@fitzgerald-fii.com

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