PA3-851

antibody from Invitrogen Antibodies

Targeting: PRKG1 PGK, PKG, PKG1, PRKG1B, PRKGR1B

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA3-851

Provider

Invitrogen Antibodies

Product name

Anti-IRAG

Antibody type

Polyclonal

Antigen

Synthetic peptide corresponding to residues 93-107 of bovine IRAG.

Host

Rabbit

Vial size

100 µl

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Regulation of intracellular calcium by a signalling complex of IRAG, IP3 receptor and cGMP kinase Ibeta.

Schlossmann J, Ammendola A, Ashman K, Zong X, Huber A, Neubauer G, Wang GX, Allescher HD, Korth M, Wilm M, Hofmann F, Ruth P
Nature 2000 Mar 9;404(6774):197-201

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of MRVI1 was achieved by transfecting Neuro-2a cells with MRVI1 specific siRNA (Silencer® select Product # 70042 and s70043). Immunofluorescence analysis was performed on Neuro-2a cells (untransfected, panel a,d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with MRVI1 specific siRNA (panel c,f). Cells were fixed, permeabilized, and labelled with MRVI1 Rabbit Polyclonal Antibody (Product # PA3-851, 1:250 dilution), followed by Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000 dilution). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Loss of signal was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to MRVI1 (green). The images were captured at 60X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of MRVI1 (IRAG) was performed using 70% confluent log phase Neuro-2a cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with MRVI1 Rabbit polyclonal antibody (Product # PA3-851) at 5 µg/mL in 0.1% BSA, incubated overnight at 4 degree Celsius and then labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and membranous localization. Panel e shows control cells with no primary antibody to assess background. The images were captured at 60X magnification.