MMP-2 and Rab11 antibodies were purchased from Abcam (rabbit ab92536 and rabbit ab3612, respectively). NUCB1, β-actin, and monoclonal anti-FLAG M2 peroxidase antibodies were obtained from Sigma-Aldrich (rabbit HPA008176 and mouse A5441 and A8592, respectively); ERGIC53 was acquired from ENZO Life Sciences (mouse ENZ-ABS300); GM130 antibody and β-integrin were purchased from BD Bioscience (mouse 610822 and mouse 610467, respectively); Rab5, Rab7, and streptavidin-HRP antibodies were from Cell Signaling (rabbit C8B1, D95F2, and 3999S, respectively); and TGN46 antibody was obtained from AbD Serotec (sheep AHP500G). MMP-14 antibody was purchased from Millipore (mouse MAB3328), and GFP antibody (rabbit sc8334) and HRP-coupled secondary antibodies (anti-rabbit IgG, anti-mouse IgG, and anti-sheep IgG) were purchased from Santa Cruz Biotechnology. Sec16A antibody (KIAA0310 polyclonal rabbit) was purchased from Biomol (now Thermo Fisher Scientific, A300-648A-M). Anti-rabbit HRP antibody used with primary macrophages was purchased from Cell Signaling (7074). The Alexa Fluor secondary antibodies used for immunofluorescence (488, 594, 633, and phalloidin-Alexa Fluor 488, different species) were purchased from Thermo Fisher Scientific.
Ergic 53
Manufactured by Enzo Life Sciences
ERGIC-53 is a type I membrane protein that functions as a cargo receptor in the early secretory pathway. It is involved in the transport of secretory cargo between the endoplasmic reticulum (ER) and the Golgi apparatus.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488, by Thermo Fisher Scientific (4 mentions)
Alexa fluor 568, by Thermo Fisher Scientific (4 mentions)
β actin, by Merck Group (2 mentions)
Gm130, by BD (2 mentions)
Gm130 antibody, by BD (1 mentions)
Tgn46 antibody, by Bio-Rad (1 mentions)
Anti mouse igg, by Santa Cruz Biotechnology (1 mentions)
Alexa fluor secondary antibody, by Thermo Fisher Scientific (1 mentions)
Anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Sec31, by BD (1 mentions)
Polyethylene glycol 1500, by Roche (1 mentions)
Flag tag (flag), by Merck Group (1 mentions)
Protein g sepharose, by GE Healthcare (1 mentions)
Calnexin, by BD (1 mentions)
Gmap210, by Merck Group (1 mentions)
α tubulin, by Merck Group (1 mentions)
Lamp1, by Developmental Studies Hybridoma Bank (1 mentions)
5 protocols using ergic 53
1
Antibody Sourcing for Cellular Imaging
2
Monoclonal Antibody Generation for ER Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A female 6-wk-old Wistar rat (CLEA Japan, Inc.) was immunized with GST-tagged Sec12 (93–239 aa) in TiterMax Gold (TiterMax USA, Inc.). Splenocytes were fused with PAI mouse myeloma cells using Polyethylene Glycol 1500 (Roche). Hybridoma supernatants were screened by indirect ELISA with ColdTF-tagged Sec12 (93–239 aa) as the antigen. Positive hybridoma lines were subcloned, grown in serum-free medium (Nihon Pharmaceutical) supplemented with HT (Life Technologies), and purified with protein G–Sepharose (GE Healthcare). Purified rabbit polyclonal antibodies against cTAGE5 (cTAGE5 CC1 [118–227 aa]; cTAGE5 C terminus [CT; 791–804 aa]), TANGO1 (1,884–1,898 aa), and collagen VII (NC2 domain) were used as described previously (Saito et al., 2009 (link), 2011 (link)). Other antibodies were purchased from the following companies: Sec16 (rabbit; Bethyl Laboratories, Inc.), KDEL (mouse; Enzo Life Sciences), β-actin (mouse; Sigma-Aldrich), EEA1 (mouse; BD), GM130 (mouse; BD), calnexin (mouse; BD), FLAG (mouse; Sigma-Aldrich), HA (rat; Roche), Sec31 (mouse; BD), and ERGIC-53 (mouse; Enzo Life Sciences).
3
Antibody Reagents for Golgi Apparatus Studies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All reagents were from Sigma-Aldrich or Merck Millipore, unless stated otherwise. Antibodies used in this study were against the following proteins: GMAP-210 (rabbit polyclonal; Sigma), GMAP-210 (mouse monoclonal; BD Biosciences), GM130 (mouse monoclonal; BD Biosciences), Myc (mouse monoclonal, 9B11; Cell Signaling Technology), Myc (rabbit polyclonal; Abcam), ERGIC53 (mouse monoclonal; Enzo Life Sciences), LAMP-1 (mouse monoclonal, H4A3; Developmental Studies Hybridoma Bank), GalNAcT2 (mouse monoclonal, UH-4; gift from Henrik Clausen), TfR (rabbit polyclonal; Millipore), GRASP65 (sheep; gift from Jon Lane), β-COP (mouse monoclonal, mAD; gift from Thomas Kreis), mannosidase II (rabbit polyclonal; Chemicon), and α-tubulin (mouse monoclonal; gift from Keith Gull). Mouse anti-p115 (4H1), rabbit anti-GM130, sheep anti-golgin-84, and sheep anti-GFP were described previously (Choudhury et al., 2009 (link); Diao et al., 2003 (link)). Sheep anti-GMAP-210 was raised against maltose binding protein fused to amino acids 1–375 of human GMAP-210 and affinity purified against the fusion protein. Fluorophore–conjugated secondary antibodies for microscopy and Western blotting were purchased from Molecular Probes and LI-COR Biosciences, respectively.
4
Immunofluorescence of Golgi Apparatus Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
One day before preparation, 5 × 10 4 fibroblasts were seeded on glass cover slips. Immunofluorescence was carried out as described [20] . Cells were stained with antibodies against ATP6AP1 (Sigma-Aldrich, rabbit anti-human, 1:300) and GM130 (BD Transduction Laboratories, mouse anti-human, 1:500) or ERGIC-53 (Enzo Life Sciences, mouse anti-human, 1:500). After incubation with fluorochrome-conjugated secondary antibodies, Alexa Fluor 488 (ThermoFisher, goat anti-rabbit, 1:500) or Alexa Fluor 568 (ThermoFisher, goat anti-mouse, 1:500), cells were mounted with Mowiol and analyzed by confocal fluorescence microscopy.
5
Immunofluorescence of Golgi Apparatus Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
One day before preparation, 5 × 10 4 fibroblasts were seeded on glass cover slips. Immunofluorescence was carried out as described [20] . Cells were stained with antibodies against ATP6AP1 (Sigma-Aldrich, rabbit anti-human, 1:300) and GM130 (BD Transduction Laboratories, mouse anti-human, 1:500) or ERGIC-53 (Enzo Life Sciences, mouse anti-human, 1:500). After incubation with fluorochrome-conjugated secondary antibodies, Alexa Fluor 488 (ThermoFisher, goat anti-rabbit, 1:500) or Alexa Fluor 568 (ThermoFisher, goat anti-mouse, 1:500), cells were mounted with Mowiol and analyzed by confocal fluorescence microscopy.
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!