Antibodies for V-ATPase Subunits Analysis

Antibodies raised against the V-ATPase ‘a’ subunit isoforms a1 and a2 were generated in our laboratory. The mouse anti-a2V neutralizing antibody against 488–510 amino acids of trans-membrane region (Antibody clone 2C1) and rabbit anti-a2NTD against N terminal domain (Antibody Clone 470) were used as described previously [25 (link), 44 (link), 60 (link)]. Anti a1 antibody was raised in rabbit against the synthetic peptides from unique regions of a1 (amino acids 73–95; RKANIPIMDTGENPEVPFPRD) by Covance (USA) and anti a3 antibody was purchased from Abnova, USA. Notch1 (antibody clone EP1238Y) and organellar markers Rab5, Pan-Cadherin and Golph4 were from Abcam. For Western Blot we used cleaved Notch1 antibody Val1744 (Cell signaling, Danvers, MA), Jagged1 (Antibody clone H114, Santa-Cruz, CA), LC3B (Abcam). β-actin (antibody clone AC-74) was purchased from Sigma Aldrich and used as the loading control. For immunohistochemistry we used Notch-1(Santa-Cruz, antibody clone C-20), and anti-a2V. For flow cytometry we used Notch1-APC (Biolegend, San Diego, CA) and FITC conjugated anti-a2V (Covance, Princeton, NJ).

Free full text: Click here

Pamarthy S., Jaiswal M.K., Kulshreshtha A., Katara G.K., Gilman-Sachs A, & Beaman K.D. (2015). The Vacuolar ATPase a2-subunit regulates Notch signaling in triple-negative breast cancer cells. Oncotarget, 6(33), 34206-34220.

Publication 2015

Notch1 Val1744 Jagged1 β-actin

Actin Amino acids Anti antibody Antibodies Antibody Atpase Cadherin Clone Fitc Flow cytometry Immunohistochemistry Isoforms Jagged1 Membrane Mouse Neutralizing antibody Organellar Peptides Rabbit Subunit Western blot

Corresponding Organization :

Other organizations : Rosalind Franklin University of Medicine and Science

Top 5 similar protocols

Variable analysis

independent variables

Anti-a2V neutralizing antibody against 488-510 amino acids of trans-membrane region (Antibody clone 2C1)
Anti-a2NTD against N terminal domain (Antibody Clone 470)
Anti-a1 antibody raised in rabbit against the synthetic peptides from unique regions of a1 (amino acids 73-95; RKANIPIMDTGENPEVPFPRD)
Anti-a3 antibody purchased from Abnova, USA

dependent variables

Notch1 expression and activation
Jagged1 expression
LC3B expression (as a marker of autophagy)

control variables

β-actin (antibody clone AC-74) as loading control for Western Blot
Rab5, Pan-Cadherin and Golph4 as organellar markers

positive controls

None specified

negative controls

None specified

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!