Ab213521

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab213521 is an antibody product offered by Abcam. It is used for research purposes. No further details are available.

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Lab products found in correlation

4 protocols using ab213521

ALK Cytotoxicity Evaluation in RAW 264.7 Cells

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Dimethyl sulfoxide (DMSO), ultra-pure E. coli K12, LPS, and MTT were purchased from Sigma. ALK (98% purity) was purchased from Sichuan Victory (ALK structure is shown in Figure 1a). ELISA kits were purchased from R&D Systems. fetal bovine serum (FBS) and dulbecco’s modified eagle medium (DMEM) were obtained from HyClone. Antibodies were purchased from Abcam (Cambridge, UK) or Proteintech (Rosemont, IL, USA), including anti-iNOS (ab210823, Abcam), anti-COX-2 (ab169782, Abcam), anti-p-P38 (ab195049, Abcam), anti-p38 (ab170099, Abcam), anti-p-ERK1/2 (ab201015, Abcam), anti-ERK1/2 (ab17942, Abcam), anti-p-JNK (ab47337, Abcam), anti-IL-6 (ab233706, Abcam), anti-IL-1β (ab254360, Abcam), anti-TNF-α (ab183218, Abcam), anti-JNK (ab213521, Abcam), anti-p-p65 (ab76302, Abcam), anti-p65 (ab16502, Abcam), anti-p-IκB-α (ab133462, Abcam), anti-IκB-α (ab32518, Abcam), and anti- glyceraldehyde-3-phosphate dehydrogenase (GADPH; ab8245, Abcam).
Figure 1.

ALK had limited cytotoxicity in RAW 264.7 cells. (a) the chemical structure of ALK was presented. (b) RAW264.7 cells were exposed to 1, 3, 5, 10 or 20 μM ALK for 24 h. The cell viability was assessed by MTT assay.

Yang J., Li J., Yang L, & Guo R. (2023). Alkannin reverses lipopolysaccharides-induced inflammatory responses by suppressing mitogen-activated protein kinase and nuclear factor kappa-B signalling. Bioengineered, 13(7-12), 14936-14946.

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Western Blot Analysis of Autophagy Markers

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Proteins were isolated with 12 % SDS–PAGE and then transferred onto homopolymers and copolymer membranes (Schleicher & Schuell, Germany). The membranes were blocked in phosphate-buffered saline (PBS) that containing 10 % nonfat dry milk and 0.5 % Tween-20 overnight. Subsequently, the membranes were incubated with primary antibodies for 2 h. The following antibodies were used: anti- LC3B (Cell Signaling Technology, Danvers, MA, USA; #2775; 1:1000), anti-p62 (Cell Signaling Technology; #5114; 1:1000), anti-HMGB2(Abcam, Cambridge Science Park, UK; ab67282; 1:1000), p-JNK (Abcam, Cambridge Science Park, UK; ab47337; 1:1000), JNK (Abcam, Cambridge Science Park, UK; ab213521; 1:1000), and anti-β-actin (Abcam, Cambridge Science Park, UK; ab8226; 1:1000). The bands were visualized using a chemiluminescence detection system (CWBIO; Beijing, China) and were normalized to that of β-actin.

Tan H.Y., Qing B., Luo X.M, & Liang H.X. (2021). Downregulation of miR-223 promotes HMGB2 expression and induces oxidative stress to activate JNK and promote autophagy in an in vitro model of acute lung injury. Journal of Inflammation (London, England), 18, 29.

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DRG Protein Expression Analysis

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Briefly, DRG tissues (L4–L6) were removed, and total protein was extracted. The
lysates were centrifuged, and the supernatants were collected. After being
denatured, the supernatant samples containing 20 μg of protein were loaded onto
gels and electrically transferred to a polyvinylidene fluoride membrane. The
membrane was incubated overnight with primary antibodies (diluted at 1:500):
rabbit anti-TRPA1 (1:500, Novus Bio, NB100-91319), anti-TNFR1 (1:500; Abcam
#ab90463), anti-IL-6R (1:500; Abcam#ab103798), anti-p-p38-MAPK (1:500; USBio,
USB#403230)/p38-MAPK (1:500; USBio, USB#403226), anti-p-JNK1(1:500; Abcam
#ab47337)/JNK1 (1:500; Abcam #ab213521). The membranes were washed and incubated
with an alkaline phosphatase conjugated antirabbit secondary antibody (1:1000).
The primary and secondary antibodies were obtained from Abcam Co. or Antibodies
online Com. The immunoreactive proteins were detected by enhanced
chemiluminescence. The bands recognized by the primary antibody were visualized
by exposure of the membrane onto an X-ray film. The membrane was stripped and
incubated with anti-β-actin to show equal loading of the protein. The film was
then scanned, and the optical density of
TRPA1/TNFR1/IL-6R//p-p38-MAPK/p38-MAPK/p-JNK1/JNK1/β-actin bands was analyzed
using the NIH Scion Image software.

Zhao D., Han D.F., Wang S.S., Lv B., Wang X, & Ma C. (2019). Roles of tumor necrosis factor-α and interleukin-6 in regulating bone cancer pain via TRPA1 signal pathway and beneficial effects of inhibition of neuro-inflammation and TRPA1. Molecular Pain, 15, 1744806919857981.

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Western Blot Analysis of DRG Proteins

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Briefly, DRG tissues (L4-L6) were removed and total protein was extracted. The lysates were centrifuged at 15, 000 x g for 15 min and the supernatants were collected. After being denatured, the supernatant samples containing 20 μg of protein were loaded onto gels and electrically transferred to a polyvinylidene fluoride membrane. The membrane was incubated overnight with primary antibodies: rabbit anti-TNFR1 (1:500, Abcam #ab90463), anti-TRPA1 (1:500, Novus Bio, NB100-91319), anti-p-p38-MAPK (1:500, USBio, USB#403230)/ p38-MAPK (1:500, USBio, USB#403226) and anti-p-JNK1(1:500, Abcam #ab47337)/ JNK1 (1:500, Abcam #ab213521). The membranes were washed and incubated (8 hours) with an alkaline phosphatase conjugated anti-rabbit secondary antibody (1:1000, Sigma, Cat#A3687). The immunoreactive proteins were detected by enhanced chemiluminescence. The bands recognized by the primary antibody were visualized by exposure of the membrane onto an x-ray film. The membrane was stripped and incubated with anti-β-actin to show equal loading of the protein. The film was then scanned and the optical density of TRPA1/TNFR1/p-p38-MAPK/ p38-MAPK/p-JNK1/JNK1/β-actin bands was analyzed using the Scion Image software.

Li C., Deng T., Shang Z., Wang D, & Xiao Y. (2018). Blocking TRPA1 and TNF-α Signal Improves Bortezomib-Induced Neuropathic Pain. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 51(5).

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