α-CD8 and rabbit α-HA were obtained from Sigma, α-BAP31, α-Actin and α-tubulin from AbCam, α-ERGIC53 from Alexis, mouse α-HA from Santa Cruz, α-EEA1 from Cell signalling and α-GM130 from BD biosciences. IRDye 800 CW and IRDye 680 RD antibodies were from LI-COR, and fluorescently conjugated secondary antibodies for microscopy were from Jackson Laboratories (Stratech Scientific). The inhibitors leupeptin (Enzo Life Sciences), pepstatin A (Sigma), Z-LLF-CHO (PSII, Calbiochem), and cycloheximide (CHX, Sigma) were used at a final concentration of 0.5 mM, 1 μg/ml, 10 μM, and 100 μg/ml respectively.
Mouse α ha
Manufactured by Santa Cruz Biotechnology
Mouse α-HA is a monoclonal antibody that recognizes the HA (Hemagglutinin) tag. The HA tag is a commonly used epitope tag for the identification and purification of recombinant proteins expressed in various systems. The Mouse α-HA antibody can be used for the detection and immunoprecipitation of HA-tagged proteins.
Automatically generated - may contain errors
Lab products found in correlation
Leupeptin, by Enzo Life Sciences (1 mentions)
α tubulin, by Abcam (1 mentions)
α actin, by Abcam (1 mentions)
α gm130, by BD (1 mentions)
Pepstatin a, by Merck Group (1 mentions)
Irdye 800cw, by LI COR (1 mentions)
Irdye 680rd, by LI COR (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Mouse α flag, by Merck Group (1 mentions)
Ezview red anti ha affinity gel, by Merck Group (1 mentions)
Mouse α v5, by Thermo Fisher Scientific (1 mentions)
Mouse α tead1, by BD (1 mentions)
Ezview red anti flag affinity gel, by Merck Group (1 mentions)
Rat α ha, by Roche (1 mentions)
Sp5 confocal microscope, by Leica (1 mentions)
4 protocols using mouse α ha
1
Antibody Sources and Inhibitor Use
2
Co-immunoprecipitation Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For co-immunoprecipitation, cell lysates were prepared in the lysis buffer (50mM Tris-HCl, pH7.5; 150mM NaCl, 1mM EDTA, 1mM EGTA, 5mM Na4P2O7, 25mM NaF, 1% Triton X-100) with protease inhibitors (cocktail) on ice for 30min. After clarification by centrifugation, the lysates were incubated for 2-6h at 4 °C with antibodies pre-bound to protein A/G agarose beads. Beads were washed four times with washing buffer (50mM Tris-HCl, pH7.5; 150mM NaCl, 1mM EDTA, 1mM EGTA, 5mM Na4P2O7, 25mM NaF, 0.5% Triton X-100), and eluted in 1x SDS loading buffer. Eluted samples were analyzed by western blotting. Antibodies used for immunoprecipitation were: mouse α-Myc (MBL International), mouse α-V5 (Thermo Fisher Scientific), mouse α-TEAD1 (BD Biosciences), EZview™ Red Anti-HA Affinity Gel (Sigma-Aldrich) and EZview™ Red Anti-Flag Affinity Gel (Sigma-Aldrich). Primary antibodies used for western blot were: rabbit α-E2F1 (X. Bi, 1:2000), mouse α-Flag (Sigma-Aldrich, 1:10000), rabbit α-Yki (L. Zhang, 1:5000), mouse α-TEAD1 (BD Biosciences, 1:500), mouse α-HA (Santa Cruz Biotechnology, 1:1000), mouse α-V5 (Thermo Fisher Scientific, 1:20000), mouse α-Myc (MBL International, 1:2000), rabbit α-hE2F1 (CST, 1:1000), mouse α-GAPDH (CWBiotech, 1:1000) and mouse α-β-actin (Proteintech Group, 1:5000).
3
Lipophorin Visualization in Drosophila
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used in this work: rabbit α-ApoLTPI and α-ApoLTP II [4 (link)], rabbit α-LpFL and rabbit α-LpII [5 (link)], rat α-HA (Roche), mouse α-HA (Santa Cruz Biotechnology) and mouse α-Myc (DSHB). Immunostaining of imaginal discs and ovaries as well as immunostaining of extracellular proteins was performed as described [13 (link)]. To examine the stability of lipophorin association to Lpr2E in vivo (Fig 7 ), wing imaginal discs were dissected in Sf-900 II SFM culture media (gibco) at 4°C and incubated in the same media for 30 minutes or 60 minutes, also at 4°C. They were subsequently fixed and processed following standard protocols. Lipids were visualized by Nile red or by oil red O stains. For Nile red, fixed imaginal discs or ovaries were incubated with 0.002% Nile red dye diluted in PBS and 0.3% triton X-100 for 60 minutes and washed for 10 minutes in the same buffer without the dye. For oil red O stain, fixed imaginal discs were incubated in a 0.5% solution of oil red O in propylene glycol at 60°C for one hour and then washed twice in 85% propylene glycol and three times in PBS, essentially as described in [56 (link)].
4
Immunostaining of Dissected Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We performed immunostaining as previously described99 (link), by fixing dissected tissues in PBS containing 4% formaldehyde and incubating with the following primary antibodies: mouse α-HA (1:1000, Santa Cruz), and guinea pig α-Mettl3 (1:2000, gift of Cintia Hongay, Clarkson University). Alexa Fluor-488, and -568 secondary antibodies were from Molecular Probes and used at 1:1000. Tissues were mounted in Vectashield mounting buffer with DAPI (Vector Laboratories). Images were captured with a Leica SP5 confocal microscope; endogenous GFP signals were monitored.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!