PA1-725

antibody from Invitrogen Antibodies

Targeting: GNB1

Western blot (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-725

Provider

Invitrogen Antibodies

Product name

Gnb1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA1-725 detects transducin beta (T beta) from sheep, bovine, mouse and drosophila. PA1-725 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~37 kDa protein representing Tr beta from sheep and bovine retinal and optic nerve extracts as well as mouse eye samples and drosophila. The PA1-725 immunizing peptide corresponds to amino acid residues 8-25 from human Tr beta. This sequence is completely conserved between human, amphibian, bovine and rat. PA1-725 immunizing peptide (Cat. # PEP-154) is available for use in neutralization and control experiments.

Reactivity

Human, Mouse, Rat, Bovine, Drosophila

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Phosphorylation of phosducin accelerates rod recovery from transducin translocation.

Belcastro M, Song H, Sinha S, Song C, Mathers PH, Sokolov M
Investigative ophthalmology & visual science 2012 May 1;53(6):3084-91

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RAS-converting enzyme 1-mediated endoproteolysis is required for trafficking of rod phosphodiesterase 6 to photoreceptor outer segments.

Christiansen JR, Kolandaivelu S, Bergo MO, Ramamurthy V
Proceedings of the National Academy of Sciences of the United States of America 2011 May 24;108(21):8862-6

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Mechanism for the regulation of mammalian cGMP phosphodiesterase6. 1: identification of its inhibitory subunit complexes and their roles.

Yamazaki A, Bondarenko VA, Matsuura I, Tatsumi M, Kurono S, Komori N, Matsumoto H, Hayashi F, Yamazaki RK, Usukura J
Molecular and cellular biochemistry 2010 Jun;339(1-2):215-33

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Phosducin regulates the expression of transducin betagamma subunits in rod photoreceptors and does not contribute to phototransduction adaptation.

Krispel CM, Sokolov M, Chen YM, Song H, Herrmann R, Arshavsky VY, Burns ME
The Journal of general physiology 2007 Sep;130(3):303-12

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Disruption of the gene encoding the beta1-subunit of transducin in the Rd4/+ mouse.

Kitamura E, Danciger M, Yamashita C, Rao NP, Nusinowitz S, Chang B, Farber DB
Investigative ophthalmology & visual science 2006 Apr;47(4):1293-301

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Phototransduction in transgenic mice after targeted deletion of the rod transducin alpha -subunit.

Calvert PD, Krasnoperova NV, Lyubarsky AL, Isayama T, Nicoló M, Kosaras B, Wong G, Gannon KS, Margolskee RF, Sidman RL, Pugh EN Jr, Makino CL, Lem J
Proceedings of the National Academy of Sciences of the United States of America 2000 Dec 5;97(25):13913-8

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on membrane enriched cell extracts of Jurkat (Lane 1) and tissue lysate of Rat Brain (Lane 2). The blot was probed with Anti- Gnb1 Rabbit Polyclonal Antibody (Product # PA1-725, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 37 kDa band corresponding to Gnb1 was observed across all lysates tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of MCF7 (Lane 1), Jurkat (Lane 2), MADA-MB-231 (Lane 3), T-47D (Lane 4) and tissue extract of Rat Brain (Lane 5). The blot was probed with Anti-Gnb1 Polyclonal Antibody (Product # PA1-725, 1 µg/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 37 kDa band corresponding to Gnb1 was observed across cell lines and tissue tested. Along with the band of interest a non-specific band at ~110 kDa was also observed.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Gnb1 was performed by loading 30 ug of HeLa (Lane 1), HeLa - Gnb1 knockout (Lane 2) Edigene lysate. The blot was probed with Anti-Gnb1 Polyclonal Antibody (Product # PA1-725, 1 µg/ml) Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to Gnb1.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5. Quantitative Western blotting of Pd-/- retinal extracts reveals decrease in transducin subunits expression. (A) Whole retinal extracts containing 100 fmol rhodopsin from WT and Pd-/- mice were separated on 10-20% polyacrylamide gels alongside 25, 50, 75, 100, 150 fmol purified transducin standard (1-5), and probed in Western blot analysis with antibodies against rod transducin subunits. (B) Fluorescence of the corresponding transducin standards bands from the top panel were plotted against the amounts of transducin in samples 1-5 (open circles), and fitted with sigmoid curves. Triangles represent the fluorescence of transducin bands from retina extracts. The number of determinations for each subunit is indicated in Table II .