Anti-A1AR antibody (ab82477), anti-active caspase-3 (ab2302), Chk pAb to albumin (ab106582), Rb pAb to MAPKAP2 (ab131531), Rb pAb to phosphor MAPKAP2 (ab63378), Ms mAb to Hsp27 (ab2790), Rb pAb to phosphor Hsp27 (ab5594), Rb mAb to NeuN (ab177487), anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody (ab201015), anti-ERK1 + ERK2 antibody (ab17942), anti-JNK1 (phospho T183) antibody (ab47337), anti-JNK1 antibody (ab110724), anti-GFAP antibody (ab10062), and Ms mAb to NeuN (ab104224) were purchased from abcam. Anti-A2aAR antibody (sc-32261), anti-A2bAR antibody (sc-28996), anti-A3AR antibody (sc-13938), β-actin (sc-47778), and GAPDH (sc-365062) were purchased from Santa Cruz. Rb pAb to P38, phosphor P38 was purchased from Cell Signaling.
Ab110724
Manufactured by Abcam
Sourced in United States
Ab110724 is a mouse monoclonal antibody that recognizes the Histidine (His) tag. It is designed for use in various immunological techniques to detect and purify His-tagged recombinant proteins.
Automatically generated - may contain errors
Lab products found in correlation
Ab92336, by Abcam (2 mentions)
Ab8448, by Abcam (2 mentions)
Ab181602, by Abcam (2 mentions)
P gsk 3β 5558, by Cell Signaling Technology (2 mentions)
Gsk 3β 12456, by Cell Signaling Technology (2 mentions)
Nbp2 36446ss, by Novus Biologicals (2 mentions)
Total protein extraction kit, by Keygen Biotech (2 mentions)
Ab32572, by Abcam (2 mentions)
Enhanced chemiluminescence detection system, by Bio-Rad (2 mentions)
Ab82477, by Abcam (1 mentions)
Ab106582, by Abcam (1 mentions)
Ab131531, by Abcam (1 mentions)
Ab2790, by Abcam (1 mentions)
Ab5594, by Abcam (1 mentions)
Ab10062, by Abcam (1 mentions)
β actin sc 47778, by Santa Cruz Biotechnology (1 mentions)
Gapdh sc 365062, by Santa Cruz Biotechnology (1 mentions)
Ab2302, by Abcam (1 mentions)
Ab276134, by Abcam (1 mentions)
Ab18197, by Abcam (1 mentions)
6 protocols using ab110724
1
Comprehensive Antibody Analysis of Cell Signaling
2
Western Blot Analysis of Key Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was used to detect the levels of JKAMP, JNK1, GSK-3β, p-GSK-3β, β-catenin, RUNX2, and OPN. Total protein was isolated from cells using a total protein extraction kit (Keygen Biotech, Nanjing, China). The protein samples were mixed with loading buffer, boiled for 5 min, separated by SDS-PAGE, and transferred to polyvinylidene fluoride membranes [29 (link), 30 (link)]. The membranes were blocked with 5% dry skim milk in Tris-buffered saline with 0.05% (v/v) Tween-20 (TBST) for 1 h and then incubated with primary antibodies against JKAMP (NBP2-36446SS) (Novus, Littleton, USA), GAPDH (ab181602), JNK1 (ab110724), β-Catenin (ab32572), RUNX2 (ab92336) and OPN (ab8448) (Abcam, Cambridge, UK), GSK-3β (12456), or p-GSK-3β (5558) (Cell Signaling Technology, Danvers, USA) at 4 °C overnight. The membrane was washed three times with TBST and then incubated with a secondary labeled anti-rabbit or anti-mouse antibody (1:3000) for 1 h. The membrane was then washed three times with TBST and developed using an enhanced chemiluminescence detection system (Bio-Rad, Hercules, USA).
3
Western Blot Analysis of Protein Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein samples (30 µg) from HMCs were loaded onto the 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and blotted on polyvinylidene difluoride (PVDF) membrane (Millipore, Billerica, MA, USA). PBS tween-20 (PBST; Sangon Biotech) solution containing 5% skimmed milk was utilized to block the non-specific sites of the membrane. After that, the membrane was labeled with diluted primary antibodies of proliferating cell nuclear antigen (PCNA; ab18197; 1:8000; Abcam, Cambridge, MA, USA), CyclinD1 (ab16663; 1:10,000; Abcam), TGF-β1 (T3176; 1:8000; Sigma), fibronectin (FN; ab2413; 1:5000; Abcam), collagen 4 (Col-4; C1926; 1:5000; Sigma), TIMP3 (ab276134; 1:5000; Abcam), Jun N-terminal Kinase (JNK; ab110724; 1:8000; Abcam), phosphorylated JNK (p-JNK; T183; 1:3000; ab47337) and GAPDH (ab9485; 1:20,000; Abcam). After labeling with horseradish-peroxidase (HRP)-conjugated secondary antibody (1:5000; Abcam), protein bands were visualized using several films and the Super Signal West Pico Chemiluminescent Substrate Kit (Pierce, Rockford, IL, USA). Quantification of protein bands was performed using the Image Lab analysis software (Bio-Rad, Hercules, CA, USA), and the intensities of protein bands were normalized to GAPDH.
4
Protein Expression and Signaling Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total cellular protein was extracted from FTC-133 or 8505C cells treated with increasing concentrations of GB1107 or TD139 (0, 10, and 100 μM) for 24 h. Equal amounts of protein were electrophoresed in 10% polyacrylamide gels and transferred to the nitrocellulose membrane. The membrane was blocked in 5% skim milk, incubated with the primary antibody at 4°C overnight, and then treated with secondary antibody at 37°C for 1 h [14 (link)]. The primary antibodies used for western blotting were purchased from Cell Signaling Technology, Danvers, MA, USA, unless otherwise specified: galectin-3 (1 : 1000 dilution; MABT51; Sigma-Aldrich), cyclin D1 (1 : 700; #2978), cleaved caspase-3 (1 : 700; #9661), PARP1 (1 : 700; ab32138; Abcam), cleaved PARP1 (1 : 700; ab32064; Abcam), phospho-p44/42 MAPK (ERK1/2) Thr202/Tyr204 (1 : 700; #9101), ERK1/2 (1 : 700; #9102), phospho-p38 MAPK Thr180/Tyr182 (1 : 700; #9216), p38 (1 : 700; #9212), phospho-JNK1 Thr183/Tyr185 (1 : 700; ab215208; Abcam), JNK1 (1 : 700; ab110724; Abcam), phospho-AKT Ser473 (1 : 700; #9271), AKT (1 : 700; #4691), β-catenin (1 : 700; #9562), MMP2 (1 : 500; MAB3308; Sigma-Aldrich), and β-actin (1 : 5000; A5441; Sigma-Aldrich).
5
Immunohistochemical Analysis of JNK1/2 in PDAC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Expression of JNK1 and -2 was determined in specimens of patients with pancreatic ductal adenocarcinoma, who underwent tumor resection at our department. Specific recombinant rabbit monoclonal antibodies were used for JNK1 (ab110724, Abcam) and JNK2 (ab76125, Abcam). Immunohistochemistry was performed using formalin-fixed and paraffin-embedded 5 µm sections in dilution 1:250 in combination with a secondary biotinylated goat anti-rabbit antibody and a Vectastain Elite ABC kit (Vector Lab, Burlingame, CA) according to the protocol of the manufacturer. For immunohistochemical analysis of orthotopic tumors, the following antibodies were used according to the protocols of the manufacturers: Ki67 (clone SP6, Thermo Fischer Scientific, Fremont, CA), p-c-Jun (54B3, Cell Signaling Technologies), MMP-2 (SAB2108458, Sigma-Aldrich, St. Louis, MO), MMP-9 (ITA1401, G-Biosciences, St. Louis, MO), Vimentin (D21H3, Cell Signaling Technologies). All samples were collected immediately during the operation with prior obtained informed consent from all patients.
6
Protein Expression Analysis by Western Blotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was used to detect the levels of JKAMP, JNK1, GSK-3β, p-GSK-3β, β-catenin, RUNX2, and OPN. Total protein was isolated from cells using a total protein extraction kit (Keygen Biotech, Nanjing, China). The protein samples were mixed with loading buffer, boiled for 5 minutes, separated by SDS-PAGE, and transferred to polyvinylidene fluoride membranes [27, 28] . The membranes were blocked with 5% dry skim milk in Tris-buffered saline with 0.05% (v/v) Tween-20
(TBST) for 1 hour and then incubated with primary antibodies against JKAMP (NBP2-36446SS) (Novus, Littleton, USA), GAPDH (ab181602), JNK1 (ab110724), β-Catenin (ab32572), RUNX2 (ab92336) and OPN (ab8448) (Abcam, Cambridge, UK), GSK-3β (12456), or p-GSK-3β (5558) (Cell Signaling Technology, Danvers, USA) at 4°C overnight. The membrane was washed three times with TBST and then incubated with a secondary labelled anti-rabbit or anti-mouse antibody (1:3000) for 1 hour. The membrane was then washed three times with TBST and developed using an enhanced chemiluminescence detection system (Bio-Rad, Hercules, USA).
(TBST) for 1 hour and then incubated with primary antibodies against JKAMP (NBP2-36446SS) (Novus, Littleton, USA), GAPDH (ab181602), JNK1 (ab110724), β-Catenin (ab32572), RUNX2 (ab92336) and OPN (ab8448) (Abcam, Cambridge, UK), GSK-3β (12456), or p-GSK-3β (5558) (Cell Signaling Technology, Danvers, USA) at 4°C overnight. The membrane was washed three times with TBST and then incubated with a secondary labelled anti-rabbit or anti-mouse antibody (1:3000) for 1 hour. The membrane was then washed three times with TBST and developed using an enhanced chemiluminescence detection system (Bio-Rad, Hercules, USA).
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!