710100

antibody from Invitrogen Antibodies

Targeting: GSK3B

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Data presented on provider website)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

710100

Provider

Invitrogen Antibodies

Product name

Anti-Phospho-GSK3B (Ser9) Antibody (9HCLC), ABfinity™ Rabbit Oligoclonal

Antibody type

Recombinant

Antigen

Synthetic peptide

Description

This antibody is predicted to react with non-human primate, mouse and rat based on sequence homology. ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale and purified with Protein A. ABfinity™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds—the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody—recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies—an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production. Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

9HCLC

Vial size

100 µg

Concentration

0.5 mg/ml

Storage

Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C

References

Submitted references

Beta-amyloid 1-42 monomers, but not oligomers, produce PHF-like conformation of Tau protein.

Manassero G, Guglielmotto M, Zamfir R, Borghi R, Colombo L, Salmona M, Perry G, Odetti P, Arancio O, Tamagno E, Tabaton M
Aging cell 2016 Oct;15(5):914-23

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of GSK-3-beta (pS9) was performed by loading 20 µg of untreated HeLa (lane 1) and HeLa treated with Insulin (100 ng/mL for 15 minutes) lysates (lane 2) using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. GSK-3-beta (pS9) was detected at ~47 kDa using ABfinity™ GSK-3-beta (pS9) recombinant rabbit oligoclonal antibody (Product # 710100) at a 1:1000 dilution in 2.5% skim milk at 4°C overnight on a rocking platform. To confirm specificity, competition was performed with the phosphopeptide (10 µg/mL) (lane 3). Detection was performed using an HRP-conjugated goat anti-rabbit secondary antibody (Product # G-21234) at a 1:5000 dilution and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Phospho-GSK3 beta pSer9 in whole cell extracts of HeLa treated with Insulin (100ng/ml, 15 min) using a Phospho-GSK3 beta pSer9 recombinant rabbit oligoclonal antibody (Product # 710100) at a dilution of 2.5 µg/mL. To confirm specificity, competition was performed by preincubation with the phosphopeptide to inhibit antibody binding (lane 1). Results show a band at ~47kDa.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin immunoprecipitation analysis of Phospho-GSK3 beta (pSer9) was performed using cross-linked chromatin from 1 x 10^6 HCT116 human colon carcinoma cells treated with serum for 0, 15, and 60 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0ul/100ul well volume of a Phospho-GSK3 beta rabbit oligoclonal antibody (Product # 710100). Chromatin aliquots from ~1 x 10^5 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1ul of eluted DNA in 2ul SYBR real-time PCR reactions containing primers to amplify the promoter region of human UBE2B, or the imprinting control region (ICR) of the human H19 locus. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representation of the human UBE2B and H19 loci are shown above the data where boxes represent exons (grey boxes = translated regions, white boxes = untranslated regions), the zigzag lines represent introns, and the straight line represents upstream sequence. Regions amplified by UBE2B and H19 primers are represented by black bars. Data courtesy of the Innovators Program.