The primary normal human FLSs and rheumatoid arthritis (RA) FLSs were purchased from Cell Applications (San Diego, CA, USA). Cells were cultured in DMEM (Thermo Fisher Scientific, Inc., Carlsbad, CA, USA) containing 10% FBS (Bio‐Rad, Hercules, CA, USA), 100 units/ml penicillin, and 100 μg/ml streptomycin (Thermo Fisher Scientific, Inc., Carlsbad, CA, USA) at 37°C in a humidified 5% CO2 atmosphere. Primary synovial cells used in this study were during the passages 3 and 6. Monoclonal rabbit anti‐β‐actin (#4970) and anti‐LDHA (#3582) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).
Anti ldha 3582
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-LDHA (#3582) is a primary antibody that specifically recognizes the lactate dehydrogenase A (LDHA) protein. LDHA is an enzyme involved in the conversion of pyruvate to lactate, a key step in glycolysis. This antibody can be used to detect and study the expression of LDHA in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Ra fls, by Cell Applications (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Bio-Rad (1 mentions)
Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions)
Anti actin, by Merck Group (1 mentions)
Anti mct4 sc 50329, by Santa Cruz Biotechnology (1 mentions)
Anti glut1 ab115730, by Abcam (1 mentions)
Citrate antigen retrieval solution, by CWBIO (1 mentions)
Anti acetyl histone h4, by Merck Group (1 mentions)
Anti gapdh, by Santa Cruz Biotechnology (1 mentions)
Anti lamin a, by Santa Cruz Biotechnology (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Anti cytochrome c, by Santa Cruz Biotechnology (1 mentions)
Anti histone h4, by Abcam (1 mentions)
4 protocols using anti ldha 3582
1
Comparative Analysis of FLS in RA
2
Protein Expression Analysis of LNCaP Cells Treated with Aneustat
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LNCaP cells were treated with vehicle control and Aneustat for 48 hr; whole cell protein extracts were resolved on SDS‐PAGE using procedures previously reported.8 Proteins were then transferred to PVDF membranes. After blocking for 1 hr at room temperature in 5% milk in TBS/0.1% Tween‐20, membranes were incubated overnight at 4°C with appropriate primary antibodies. Following incubation with secondary antibody, immunoreactive proteins were visualized with a SuperSignal™ West Femto Maximum Sensitivity Substrate (Thermo Scientific; Rockford, IL). The following antibodies were used: anti‐GLUT1 (ab115730, rabbit monoclonal antibody, Abcam; Toronto, Canada); anti‐LDHA (3582, rabbit monoclonal antibody, Cell Signaling Technology; Beverly, MA); anti‐MCT4 (sc‐50329, rabbit polyclonal antibody, Santa Cruz; Mississauga, Canada); anti‐actin (A2066, rabbit polyclonal antibody, Sigma). Actin was used to monitor the amounts of samples applied.
3
Immunohistochemical Analysis of LDHA-K222 Succinylation
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The tissues were fixed in 10% formalin, and then embedded in paraffin. The specific steps were performed as previously described [13 (link)]. Rabbit polyclonal antibodies against the human LDHA K222-succinylated peptide, PDLGTDK (suc) DKEQWK, were generated from two rabbits (#1 and #2) at ChinaPeptides Co., Ltd. (Shanghai, China). For immunohistochemical staining, 5-μm-thick serial sections were used to prepare the slides. Antigen retrieval was performed with 10 mM citrate antigen retrieval solution (CW Biotech, Beijing, China). Anti-LDHA (#3582S, Cell Signaling Technology, Danvers, MA, USA) and anti-LDHA-K222suc antibodies was used at a dilution of 1:400. The immunostaining index was based on the staining intensity and percentages of positively stained tumor cells. The intensity was scored between 0 and 3: 0 (no staining), 1 (weak staining), 2 (moderate staining), and 3 (strong staining). The percentages of positively stained tumor cells were defined as 0 (< 10%), 1 (10 –30%), 2 (31 –50%), and 3(> 50% of positive cells) [25 (link)]. The staining index was then calculated as the score of staining intensity multiplied by the percentage of positively stained tumor cells. Tumor stage was classified according to the seventh edition of the UICC/AJCC TNM staging system.
4
Western Blot Protein Detection
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Western blot analysis was performed according to a standard protocol. The following antibodies were used to detect each target protein: anti-LDHA (3582 S, Cell Signaling, Danvers, MA, USA), anti-Lamin A (sc-71481), anti-GAPDH (sc-31915), anti-Cytochrome C (sc-13560), anti-β-actin (sc-47778, Santa Cruz, Dallas, TX, USA), anti-Histone H4 (ab10158, Abcam, Cambridge, UK), anti-acetyl-histone H4 (#06-866, Merck, Kenilworth, NJ, USA), anti-mouse (sc-2005), anti-rabbit (sc-2004), and anti-goat IgG-HRP (sc-2922, Santa Cruz, Dallas, TX, USA) antibodies.
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