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You are here:Home » Antibodies » Abs to G Proteins » Anti -G Protein, i alpha 1, (Gia1)

Anti -G Protein, i alpha 1, (Gia1)

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Specifications

Clone Host Grade Applications
Monoclonal Mouse Affinity Purified E B
Catalog #G8599-04
ApplicationsSuitable for use in ELISA and Immunoblot. Other applications not tested.
Recommended DilutionImmunoblot Analysis: 2-4ug/ml detects Gia1 from a rat brain microsomal preparation. Rat brain microsomal preparation was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Gia1 (2ug/ml). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP* and a chemiluminescence detection system.
Optimal dilutions to be determined by the researcher.
*Recommended secondary antibodiesI1904-06C: IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo
I1904-06H IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo
I1904-14C: IgG, Fc (HRP) Pab Gt xMo
I1904-19C: IgG, F(abí)2 (HRP) Pab Gt xMo
Immunoblot Protocol1. Perform SDS-PAGE on a microsomal membrane preparation 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 (PBS-MLK) for 60 minutes at RT with constant agitation.
3. Incubate the nitrocellulose with 2-4ug/ml of anti-Gia1, G8599-04 diluted in freshly prepared PBS-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, 1:3000 dilution was used)* in PBS-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 10 minutes.
8. Rinse the nitrocellulose in 4-5 changes of water for 2 hours.
9. Use detection method of choice (enhanced chemiluminescence was used).
Storage and StabilityMay be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Clone TypeMonoclonal
IsotypeIgG2b
Clone No4i40
HostMouse
SourceHuman
Concentration~1mg/ml
FormSupplied as a liquid in 0.07M Tris-glycine, pH 7.4, 0.105M NaCl, containing 30% glycerol.
PurityPurified by Protein G affinity chromatography.
ImmunogenA synthetic peptide (CAGAAEEGFMTAELA) corresponding to amino acids 110-123 of Gia1 (KLH).
SpecificityRecognizes Gia1, Mr 41kD. Approximately 30% crossreactivity with Gia3. Species Crossreactivity: Broad species crossreactivity expected due to the highly conserved nature of Gia1.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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