GNB2L1 antibody (AA 113-317)

Catalog No. ABIN967796

The Mouse Monoclonal anti-GNB2L1 antibody has been validated for WB, IHC, IP and BI. It is suitable to detect GNB2L1 in samples from Human, Mouse, Rat, Dog, Cow, Chicken and Frog. There are 5+ publications available.

Quick Overview for GNB2L1 antibody (AA 113-317) (ABIN967796)

Target: GNB2L1 (Guanine Nucleotide Binding Protein (G Protein), beta Polypeptide 2-Like 1 (GNB2L1))

Reactivity: Human, Mouse, Rat, Dog, Cow, Chicken, Frog

Host: Mouse

Clonality: Monoclonal

Conjugate: This GNB2L1 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), BioImaging (BI)

Clone: 20-RACK1

Product Details anti-GNB2L1 Antibody

Binding Specificity: AA 113-317

Cross-Reactivity: Human, Cow (Bovine), Chicken, Dog (Canine), Frog, Mouse (Murine)

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.
7. All other brands are trademarks of their respective owners.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Rat RACK1 aa 113-317

Isotype: IgM

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN968537

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-GNB2L1 antibody (ABIN967796)

Birikh, Sklan, Shoham, Soreq: "Interaction of "readthrough" acetylcholinesterase with RACK1 and PKCbeta II correlates with intensified fear-induced conflict behavior." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, Issue 1, pp. 283-8, (2003) (PubMed).

Liedtke, Yun, Kyle, Wang: "Protein kinase C epsilon-dependent regulation of cystic fibrosis transmembrane regulator involves binding to a receptor for activated C kinase (RACK1) and RACK1 binding to Na+/H+ exchange regulatory factor." in: The Journal of biological chemistry, Vol. 277, Issue 25, pp. 22925-33, (2002) (PubMed).

Chang, Chiang, Cartwright: "The interaction of Src and RACK1 is enhanced by activation of protein kinase C and tyrosine phosphorylation of RACK1." in: The Journal of biological chemistry, Vol. 276, Issue 23, pp. 20346-56, (2001) (PubMed).

Smart, Ying, Anderson: "Hormonal regulation of caveolae internalization." in: The Journal of cell biology, Vol. 131, Issue 4, pp. 929-38, (1996) (PubMed).

Ron, Chen, Caldwell, Jamieson, Orr, Mochly-Rosen: "Cloning of an intracellular receptor for protein kinase C: a homolog of the beta subunit of G proteins." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 91, Issue 3, pp. 839-43, (1994) (PubMed).

Target Details for GNB2L1

Target: GNB2L1 (Guanine Nucleotide Binding Protein (G Protein), beta Polypeptide 2-Like 1 (GNB2L1))

Alternative Name: RACK1

Background: Several proteins which specifically bind to PKC have been classified as RACKs (receptors for activated C-kinase). RACK1 was cloned from a rat brain cDNA expression library by screening for proteins that bind PKC in the presence of phosphatidylserine, diacylglycerol, and calcium in a PKC overlay assay. By sequence homology, RACK1 appears to belong to a superfamily that includes the ß subunit of G proteins. All of these proteins contain five to eight internal repeat elements known as WD40 motifs, which appear to have a role in protein-protein interactions. In addition, RACK1 contains two short sequences homologous to a PKC-binding sequence identified in Annexin I and in the brain PKC inhibitor KCIP. The binding of RACK1 to PKC is dose-dependent and occurs at a site on PKC that is distinct from the catalytic domain, indicating that RACK1 is not a PKC substrate.

Molecular Weight: 36 kDa

Pathways: cAMP Metabolic Process, Positive Regulation of Endopeptidase Activity