rev:    July 14, 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, 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.

ANTI-Tubulin  (see also antibodies for beta tubulin, and   beta III isoform  and   gamma)
Mouse anti-Alpha Tubulin (Multi-Species)   Paraffin Sections/Western Blot

cat# RDI-TUBULAAbm-DM       $438.00/100ug

       RDI-TUBULAabm-DM1       $688.00/200ug

Presentation: purified antibody in 10mM PBS pH7.4 with 0.2% BSA and 15mM sodium azide (200ug/ml)

-also available azide/BSA free   cat#RDI-TUBULAabm-DMX $750.00/200ug



Clone ref: DM1A

Isotype: mIgG1a

Immunogen: Native chick brain microtubules

Hybridoma: fusion between BALB/c57 splenocytes and myeloma NS-1 cells.

Species Reactivity: Human, Cow, Pig, Rat,Mouse, Guinea pig, Gerbil, Frog, Chicken, Yeast, and Fungi.

                                Others-not tested.



Use: -western blot (1ug/ml for 2 hours RT)

         -histochemsitry: frozen sections and formalin fixed paraffin sections (no special pretreatment required)

          -electron microscopy & immunofluorescence

Positive Control: LS174T, MAD109 cells. Skin or lung

Localization: cytoplasmic

Specificity: Recognizes a protein of 57kDa, identified as tubulin-alpha. This Mab is highly specific to alpha tubulin and shows no cross-reaction with Beta tubulin . Its epitope localizes between aa 426-450; aa 426-430 are critical for reactivity.2 . This Mab causes the 10-nm filaments to collapse into large lateral aggregates collecting in the cell periphery or tight juxtanuclear caps. It does not block microtubule assembly. Mabdoes not inhibit polymerization or depolymerization of platelet tubulin in vitro. It blocks (by 70-80%) the ability of tubulin dimers (with GppNHp bound) to promote a stable inhibition of adenylyl cyclase.

Key References:

1. Blose SH, et. al. Journal of Cell Biology, 1984, 98(3):847-58.

2. Breitling F, et. al. Journal of Molecular Biology, 1986, 189(2):367-70.

3. Biochim Biophys Acta, 1989, 1012(1):46-56.

4. Biochemistry, 1993, 32(18):4955-61.

For Research Use Only

Sheep anti-Tubulin (affinity purified)

Catalog#: RDI-TUBULabSX      $625.00/1mg     $500.00/mg 5+

Amount: 1 mg (lyophilized), Reconstitute with 1 ml di-H20 or PBS for a 1 mg/ml solution.

Preservative: None

Storage: 2 - 8 C, Stable for 6 months after reconstitution.

Production: The tubulin antigen was isolated from bovine brain origin. The antiserum was produced in sheep by repeated immunizations with the tubulin antigen. The antiserum was affinity purified over 6S tubulin Sepharose. Each vial contains 1 mg IgG + 2 mg bovine serum albumin.

Performance: Useful in the immunohistochemical and immunofluorescent detection of alpha and beta tubulin in tissue sections and cultured cells. Reacts with both interphase and mutotic tubulins and with soluble monomers (6S). The antibody cross reacts with tubulin is human, monkey, hamster and rat. Inhibits tubulin polymerization (assembly) in vitro. A dilution 1/10 is suggested for experiments.

For Research Use Only

DATA SHEET: (mouse anti-Tubulin A & B)


Catalog#: RDI-tubulinabm  $438.00

Larger sizes quoted on request

Package Size: lyophilized, reconstitute to 0.1ml with sterile PBS buffer and 0.1%NaN3,

lot#


Ig Isotype: mouse IgM (ascites)

Antigen: Calf Brain tubulin, highly purified

Dilution: approx 1:20-1:100 (optimize for each use)

Reactivity: reacts with human, mouse, rat and hamster tubulin. In immunoblotting, recognizes only tubulin. On immunofluorescence, reacts with both alpha and beta microtubules but not with   microfilaments or intermediate filaments.

Storage: Store at -20 DEG C Upon receipt Avoid repeated   freeze thaw cycles.

Ref# Cell 12:561, 1977

Sample Staining Protocol:
1. Prepare fresh DTSP (dithiobis succinimidyl propionate; from Pierce Chemical Co.) Weigh 5mg, add 0.25ml DMSO and just before use add 24.75 ml warm PBS.

2. Cell grown on cover slips are fixed in DTSP 5min/37 DEG C

3. Wash coverslip in PBS, then distilled water

4. Fix in acetone (-20 DEGC) for 2-5 min

5.  Wash in water, then PBS

6. Incubate with 2% BSA for 15min/37 DEG C.

7. Incubate with tubulin antibodies for 60min/37 DEG C.

8. Wash in PBS

9. Incubate with rhodamine-conjugated goat anti-mouse IgM for 60 min 37 DEg C.

10. Wash in PBS, then water before mouunting slide.


Precautions: For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics. Sodium azide may form explosive compounds in presence of heavy metals or under acidic conditions. Flush drains with copious amounts of water to prevent buildup of explosive compounds.

band approx 52-54kda with HElA, NIH/3T3 or A-431 whole cell lysates

RDI Divison of researchd Industries Intl

San Jose, 95123 CA Snell ave 658

USA

or 408-780-0908

EMAIL:margaret@cellular-research.com

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