Western blotting analysis was carried out as previously described [25 (link)]. The following antibodies were used: anti-Bcl-2 (#ab182858), anti-Bax (#ab182733), anti-Bcl-xl (#ab32370), anti-p-P65 (#ab76302), anti-P65 (#ab32536), anti-p-IκBα (#ab133462), anti-IκBα (#ab32518) (Abcam Cambridge, MA, USA), anti-FXR (#sc-25309), anti-TLR4 (#sc-293072) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-p-ERK1/2 (#4370), anti-ERK1/2 (#4695), anti-p-P38 (#4511), anti-P38 (#8690), anti-p-JNK1/2 (#4668), anti-JNK1/2 (#9252), anti-CCL2 (#66272-1-Ig), and anti-β-actin (#66009-1-Ig) (Proteintech, Wuhan, China).
Anti ccl2
Manufactured by Proteintech
Sourced in China
Anti-CCL2 is an antibody product developed by Proteintech for use in research applications. It is designed to detect and bind to the CCL2 protein, also known as Monocyte Chemoattractant Protein-1 (MCP-1), which is a chemokine involved in the recruitment and activation of monocytes and other immune cells. The antibody can be used for various research purposes, such as immunoassays, immunohistochemistry, and western blotting, to study the role of CCL2 in biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (3 mentions)
Ecl reagent, by Beyotime (2 mentions)
Anti gapdh, by Merck Group (2 mentions)
Anti e cadherin, by Cell Signaling Technology (2 mentions)
Anti hpv16 e6 hpv18 e6, by Abcam (2 mentions)
Anti taz, by Proteintech (2 mentions)
Anti vimentin, by Abcam (2 mentions)
Chemidoc xrs system, by Bio-Rad (2 mentions)
Anti hpv16 e7, by Abcam (2 mentions)
Anti yap, by Proteintech (2 mentions)
Anti bax, by Abcam (1 mentions)
Anti bcl 2, by Abcam (1 mentions)
Anti p65, by Abcam (1 mentions)
Anti bcl xl, by Abcam (1 mentions)
Anti β actin, by Proteintech (1 mentions)
Anti p p65, by Abcam (1 mentions)
Anti p iκbα, by Abcam (1 mentions)
Anti fxr, by Santa Cruz Biotechnology (1 mentions)
Anti p erk1 2, by Proteintech (1 mentions)
Anti p p38, by Proteintech (1 mentions)
6 protocols using anti ccl2
1
Western Blotting Analysis of Apoptosis and Inflammation Signaling
2
Comprehensive Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot was performed as previously described (35 (link)). Primary antibodies used were: anti-TNFAIP6 (1:1000, Proteintech, China), anti-TLR6 (1:1000, ABclonal, China), anti-FAS (1:3000, Proteintech, China), anti-CCL3 (1:1000, Proteintech, China), anti-ICAM-1 (1:3000, Proteintech, China), anti-CCL2 (1:3000, Proteintech, China), anti-CXCR4 (1:3000, Proteintech, China), anti-VEGFA (1:2000, Proteintech, China), anti-β-Tubulin (1:20000, Proteintech, China) and anti-GAPDH (1:2000, Servicebio, China).
3
Immunohistochemical Analysis of Inflammatory Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tissue sections were processed with an Envision Flex+kit (DAKO), blocking endogenous peroxidase activity for 5 min, and then incubated with primary antibodies. The reaction was visualized by Envision Flex + horseradish peroxidase for 20 min and then with diaminobenzidine for 10 min. Sections were counterstained with Mayer’s hematoxylin for 5 min. Primary antibodies used for the study were: anti-CD4 (Dako, Santa Clara, CA, USA, ready to use, or Abcam (Cambridge, UK, ab846), 1:50 for 20 min), anti-IBA1 (Wako, Osaka, Japan, 1:300 for 30 min), anti-GFAP (Dako, ready to use, for 20 min), anti-TNFa (ab6671 from Abcam, Cambridge, UK, 1:2000 for 45 min), anti-CCl2 (66272, Proteintech, Rosemont, IL, USA, 1:1000), anti-CCl20 (ab9829, Abcam; 1:200 overnight), anti-CX3CL1 (14-7986 from Invitrogen, Waltham, MA, USA, 1:200 overnight).
4
Comprehensive Protein Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The whole proteins were extracted using cell lysis buffer. Then, SDS-PAGE was used to separate proteins with different sizes. After that, protein was transferred to a PVDF membrane (Millipore) and target protein was immunoblotted with specific primary antibody. Primary antibodies were listed as follows: anti-E-cadherin (1:5000, mouse anti-human, Cell Signaling Technology Inc.), anti-Vimentin (1:2000, mouse anti-human, Abcam), anti-HPV16 E6 + HPV18 E6 (1:1000, mouse anti-human papillomavirus, Abcam), anti-HPV16 E7 (1:1000, mouse anti-human papillomavirus, Abcam), anti-YAP (1:2000, mouse anti-human, Proteintech), anti-TAZ (1:2000, mouse anti-human, Proteintech), anti-CCL2 (1:2000, mouse anti-human, Proteintech), anti-flag-YAP (1:1000, mouse, Abbkine), anti-GAPDH (1:2000, rabbit anti-human, Sigma-Aldrich). After incubating goat anti-rabbit or goat anti-mouse secondary antibody (MultiSciences), immunoblots were visualized using the chemiluminescence (ECL) reagent (Beyotime Biotechnology) and ChemiDoc XRS+ System (Bio-Rad).
5
Western Blot Analysis of Extracellular Matrix Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins in cells were extracted using RIPA lysis buffer (Beijing Solarbio Science & Technology Co. Ltd., Beijing, China). 8ul denatured sample proteins and 4 µL markers were loaded into 12% Sodium Dodecyl Sulfate (SDS) gel and separated by SDS-polyacrylamide gel electrophoresis. After that, we used electrotransfection to transfer the proteins to PVDF membranes (Millipore Corp, billerica, MA, USA) at 200mA for 1h. Then, Transferred PVDF membrane was blocked in incubation liquid (TBST+dried skim milk) for more than 2 h. Next, we incubated the PVDF membrane with primary antibody (anti-TnC, anti-fibronectin, anti-nidogen, anti-emilin, anti-VEGF-A, anti-VEGF-B, Abcam, USA; anti-CCL2, Proteintech, China) at 4 °C overnight. After washing with TBSP and the membrane was incubated with secondary antibodies (IgG-HPR of goat anti-rabbit and goat anti-mouse, Beijing Zhongshan Jinqiao, China) for 2 h at room temperature. Finally, we scanned the blots using a gel imaging system and used software to calculate the gray value.
6
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The whole proteins were extracted using cell lysis buffer. Then, SDS-PAGE was used to separate proteins with different sizes. After that, gel was transferred to a PVDF membrane (Millipore) and target protein was immunoblotted with speci c primary antibody. Primary antibodies were listed as follows: anti-Ecadherin (1:5000, mouse anti-human, Cell Signaling Technology Inc.), anti-Vimentin (1:2000, mouse antihuman, Abcam), anti-HPV16 E6 + HPV18 E6 (1:1000, mouse anti-human papillomavirus, Abcam), anti-HPV16 E7 (1:1000, mouse anti-human papillomavirus, Abcam), anti-YAP (1:2000, mouse anti-human, Proteintech), anti-TAZ (1:2000, mouse anti-human, Proteintech), anti-CCL2 (1:2000, mouse anti-human, Proteintech), anti-ag-YAP (1:1000, mouse, Abbkine), anti-GAPDH (1:2000, rabbit anti-human, Sigma-Aldrich). After incubating goat anti-rabbit or goat anti-mouse secondary antibody (MultiSciences), immunoblots were visualized using the chemiluminescence (ECL) reagent (Beyotime Biotechnology) and ChemiDoc XRS + System (Bio-Rad).
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!