The antibodies used for western blotting (WB), Co-IP, IHC and immunofluorescence (IF) in this study included antiSNAI2 antibody (9585, CST, USA), anti-USP7 antibody (66,514–1-lg, Proteintech, China), anti-TRIM21 antibody (12,108–1-AP, Proteintech, China), anti-CD34 antibody (11,265–1-AP, Proteintech, China), anti-Flag antibody (66,008–4-Ig, Proteintech, China), anti-Myc antibody (16,286–1-AP, Proteintech, China), anti-HA antibody (51,064–2-AP, Proteintech, China), mouse IgG (3420, CST),,protein A/G agarose beads, anti-Flag Magnetic Beads, anti-Myc Magnetic Beads, anti-HA Magnetic Beads (Beyotime, China) and MG132 (C2211, Sigma Aldrich) was dissolved in dimethyl sulfoxide.
Anti myc magnetic beads
Manufactured by Beyotime
Sourced in China
The Anti-Myc magnetic beads are a laboratory tool used for the purification and isolation of proteins tagged with the c-Myc epitope. These beads are coated with an antibody specific to the c-Myc tag, allowing for the efficient capture and recovery of the target protein from complex samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti flag magnetic beads, by Beyotime (2 mentions)
Anti ha antibody 51 064 2 ap, by Proteintech (1 mentions)
Mg132, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
Lysis buffer, by Beyotime (1 mentions)
Anti flag anti ha, by Beyotime (1 mentions)
Ip lysis buffer, by Beyotime (1 mentions)
Protein a g magnetic beads, by Beyotime (1 mentions)
4 protocols using anti myc magnetic beads
1
Western Blot and Immunoprecipitation Techniques
2
Protein Extraction and Immunoanalysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins were extracted by ristocetin‐induced platelet agglutination buffer (Solarbio), and WB, Co‐IP, and confocal microscopy were performed as previously described.
12 (link) Anti‐RTN3 primary antibody (1:2000) was self‐produced. Anti‐GAPDH primary antibody (10494‐1‐AP, 1:5000), anti‐flag primary antibody (20543‐1‐AP, 1:5000), and anti‐p62 primary antibody (18420‐1‐AP, 1:1000) were purchased from Proteintech Company (Wuhan, China). Anti‐SPHK2 primary antibody (YT4383, 1:1000) and anti‐CYP27A1 primary antibody (YT1202, 1:1000) were purchased from ImmunoWay Biotechnology Company (Suzhou, China). Anti‐myc primary antibody (sc‐40, 1:100), anti‐CERS2 primary antibody (sc‐390745, 1:100), and anti‐LC3A/B primary antibody (sc‐398822, 1:100) were purchased from Santa Cruz Biotechnology Incorporated (Santa Cruz, USA). Anti‐myc magnetic beads (Beyotime) and anti‐flag magnetic beads (Beyotime) were used for Co‐IP, and self‐produced anti‐RTN3 primary antibody (1:400) and anti‐CD31 primary antibody (66065‐2‐Ig, 1:200) were purchased from Proteintech Company; anti‐LC3 primary antibody (PM036, 1:200) were purchased from MBL International Corporation (Kyushu, Japan); anti‐CERS2 primary antibody (sc‐390745, 1:100) and DAPI (Solarbio) were used for confocal microscopy.
12 (link) Anti‐RTN3 primary antibody (1:2000) was self‐produced. Anti‐GAPDH primary antibody (10494‐1‐AP, 1:5000), anti‐flag primary antibody (20543‐1‐AP, 1:5000), and anti‐p62 primary antibody (18420‐1‐AP, 1:1000) were purchased from Proteintech Company (Wuhan, China). Anti‐SPHK2 primary antibody (YT4383, 1:1000) and anti‐CYP27A1 primary antibody (YT1202, 1:1000) were purchased from ImmunoWay Biotechnology Company (Suzhou, China). Anti‐myc primary antibody (sc‐40, 1:100), anti‐CERS2 primary antibody (sc‐390745, 1:100), and anti‐LC3A/B primary antibody (sc‐398822, 1:100) were purchased from Santa Cruz Biotechnology Incorporated (Santa Cruz, USA). Anti‐myc magnetic beads (Beyotime) and anti‐flag magnetic beads (Beyotime) were used for Co‐IP, and self‐produced anti‐RTN3 primary antibody (1:400) and anti‐CD31 primary antibody (66065‐2‐Ig, 1:200) were purchased from Proteintech Company; anti‐LC3 primary antibody (PM036, 1:200) were purchased from MBL International Corporation (Kyushu, Japan); anti‐CERS2 primary antibody (sc‐390745, 1:100) and DAPI (Solarbio) were used for confocal microscopy.
3
Ubiquitination and Interaction of FSP1 with ACSL1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HA-Ub and Myc-FSP1 expression plasmids together with Flag-ACSL1-WT or Flag-ACSL1-MT were co-transfected into HEK293T cells. Cells were treated with 20 μM MG132 44 h post-transfection for 4 h and then lysed using lysis buffer (P0013, Beyotime). The cell lysates were immunoprecipitated by anti-Myc magnetic beads (P2118, Beyotime) overnight at 4°C. The immunocomplexes were subjected to western blotting. The ubiquitin level of FSP1 was detected using an anti-HA antibody (cat:3724, CST). The expression of FSP1 and ACSL1 was detected by anti-FSP1 and anti-ACSL1 antibodies, respectively.
4
Exploring TRIM69 Protein Interactions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect the interaction of endogenous TRIM69 with PRKCD, AGS, MKN28 or MKN45 cell lysates were obtained by using IP lysis buffer (Beyotime, Shanghai, China) and incubated with 2 µg of an anti-PRKCD antibody or anti-rabbit IgG (Beyotime, Shanghai, China) overnight at 4°C. Protein A + G magnetic beads (Beyotime, Shanghai, China) were added to the cell mixtures and incubated for 4 h at 4°C. For the interaction between exogenous TRIM69 and PRKCD, AGS, MKN28, MKN45 or HEK293T cells were transfected with TRIM69 or PRKCD plasmids. After culture for 48 h, the cells were homogenized in lysis buffer for immunoprecipitation (Beyotime, Shanghai, China). Then, the cell lysates were incubated with 20 µl of anti-IgG magnetic beads (Beyotime, Shanghai, China) overnight at 4°C. The next day, the mixtures were incubated with anti-Flag, anti-HA or anti-Myc magnetic beads (Beyotime, Shanghai, China) for 4 h at 4°C. After the beads were washed four times with lysis buffer for IP, the precipitates were collected, and proteins were eluted with 1×SDS sample buffer. The samples were analyzed using western blotting.
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!