Baicalin (purity ≥ 98%) was obtained from Xi’an Kai Lai Biological Engineering Co., Ltd. (Xi’an, China). Fluoxetine hydrochloride (Flu) was purchased from Changzhou Siyao Pharmaceuticals Co., Ltd. (Changzhou, China). Lipopolysaccharide (LPS), 4,6-diamidino-2-phenylindole (DAPI), and compound C were bought from Sigma-Aldrich Co (St. Louis, USA). TAK-242 (a TLR4 antagonist) and LY294002 (a PI3K inhibitor) were products purchased from Apex Bio (Houston, USA). Poly-d -lysine was obtained from Sigma (MO, USA). Interleukin-1β (IL-1β), Interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α) enzyme-linked immunosorbent assay (ELISA) kits were supplied by Elabscience Biotechnology Co., Ltd. (Wuhan, China). The antibodies were obtained from the cited commercial sources: anti-p-PI3K (#4228), anti-PI3K (#4292), anti-p-Akt (Ser473, #9271), anti-Akt (#9272), anti-β-actin (#4967), anti-PCNA (#13110), anti-p-AMPK (#2531), and anti-AMPK (#2603) were from Cell Signaling Technology (Beverly, MA, USA); anti-PTEN (sc-7974) and anti-p-PTEN (sc-377573) were from Santa Cruz Biotechnology (Santa Cruz, CA); anti-TLR4 (AF7017), anti-p-GSK3β (AF2016), and anti-GSK3β (AF7814) were from Affinity Biosciences (Changzhou, China); and anti-FoxO1 (ab52857), anti-p-FoxO1 (Ser 256, ab131339), and goat anti-rabbit IgG H&L (Alexa Fluor® 488, ab150077) were from Abcam (Cambridge, MA, USA).
Goat anti rabbit igg h l alexa fluor 488 ab150077
Manufactured by Abcam
Sourced in United States, United Kingdom
Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077) is a secondary antibody conjugated with Alexa Fluor® 488 dye. It is designed for the detection of rabbit primary antibodies in various immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Mitotracker red, by Thermo Fisher Scientific (3 mentions)
Cell total protein extraction kits, by Beyotime (2 mentions)
Ab6671, by Abcam (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (2 mentions)
H2dcfda, by Thermo Fisher Scientific (2 mentions)
Tumor necrosis factor α tnf α enzyme linked immunosorbent assay elisa kits, by Elabscience (1 mentions)
Anti pten sc 7974, by Santa Cruz Biotechnology (1 mentions)
Tak 242, by Apexbio (1 mentions)
Anti p ampk, by Cell Signaling Technology (1 mentions)
Anti ampk, by Cell Signaling Technology (1 mentions)
Anti pi3k, by Cell Signaling Technology (1 mentions)
Anti p pi3k, by Cell Signaling Technology (1 mentions)
Anti pcna, by Cell Signaling Technology (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Compound c, by Merck Group (1 mentions)
Poly d lysine, by Merck Group (1 mentions)
Ly294002, by Apexbio (1 mentions)
P38 inhibitor sb203580, by Selleck Chemicals (1 mentions)
Jnk inhibitor sp600125, by Selleck Chemicals (1 mentions)
U0126, by Selleck Chemicals (1 mentions)
20 protocols using goat anti rabbit igg h l alexa fluor 488 ab150077
1
Baicalin Modulates TLR4/PI3K/AMPK Pathway
2
Antibody Detection and MAPK Pathway Inhibitors
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Antibodies against EGR1 (#4154), cJUN (#9165), cFOS (#2250), ERK1/2 (#9194), JNK (#9252), p38 (#9212), PARP (#9532), β-actin (#4967) were purchased from Cell Signaling Technology. Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077) was purchased from Abcam. Antisera against IBV N protein were prepared in rabbits immunized with bacterially expressed fusion proteins as previously described [27 (link),28 (link)]. MEK inhibitor U0126, p38 inhibitor SB203580 and JNK inhibitor SP600125 were purchased from Selleckchem.
3
Synthesis and Characterization of PEG-DA Hydrogel
4
Conditional Knockout Mice for Immune Cell Analysis
5
Panax notoginseng Saponins Modulate Bone Metabolism
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PNSs, the total saponins of Panax notoginseng, were purchased from KPC Xuesaitong Pharmaceutical Co., Ltd (Yunnan, China). C57BL/6J mice were purchased from SPF (Beijing) Biotechnology Co., Ltd (Beijing, China). Mouse NTXI(cross linked N-telopeptide of type I collagen) ELISA kit (E-EL-M3022) was purchased from Elabscience (Wuhan, China). Anti-osteocalcin antibody (ab93876), recombinant anti-CD31 antibody (ab182981), goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077), and goat anti-rabbit IgG H&L (HRP) (ab205718) were purchased from Abcam (USA).
6
DiD-Based Fluorescence Imaging Assay
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1,19-Dioctadecyl-3,3,39,39-tetramethylindodicarbocyanine perchlorate (DiD) was purchased from Biotium Inc. (Fremont, USA). DAPI, cell total protein extraction kits, and N-acetyl-l -cysteine (NAC) were provided by Beyotime Institute of Biotechnology (Jiangsu, China). Antibodies against CD31 (ab28364) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) were obtained from Abcam. l -Ascorbic acid (VC) was purchased from Aladdin (Shanghai, China). Malondialdehyde (MDA), superoxide dismutase (SOD), and monocyte chemotactic protein 1 (MCP-1) enzyme-linked immunosorbent assay (ELISA) kits were provided by KeyGen Biotech (Nanjing, China).
7
Antibody Characterization Protocol
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The antibodies and fluorescent dyes used in this study were as follows: anti-Flag (FUJIFILM Wako, 018-22381), normal mouse IgG (Santa Cruz, sc-2025), and calcein AM (MB5279, Meilunbio). Anti-GAPDH (14C10) rabbit mAb (2118S), LaminA/C (4C11) mouse mAb (4777S), and Myc-tag (9B11) mouse mAb (2276S) were purchased from Cell Signaling Technology. Anti-SUN2 antibody [EPR6557] (ab124916), goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077), and goat anti-mouse IgG H&L (Alexa Fluor® 555) (ab150114) were purchased from Abcam. Peroxidase AffiniPure goat anti-mouse IgG (H+L) (115-035-003) and peroxidase AffiniPure rabbit anti-goat IgG (H+L) (305-035-003) were purchased from Jackson ImmunoResearch Laboratories. F-actin monoclonal antibody (NH3) (MA1-80729), Alexa Fluor 488 Isolectin GS-IB4 conjugate (I21411), and Hoechst 33342 solution (H3570) were purchased from Invitrogen. γ-tubulin antibody (T6557) and BSI lectin-FITC (L9381) were purchased from Sigma-Aldrich. The anti-TMEM201 rabbit antibody, generated with the TMEM201 (430–638 amino acid) peptide, was made by Youke Biological Technology Co. Ltd.
8
Quantifying Tumor Cell Proliferation via Ki-67 IHC
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Immunohistochemical detection of Ki-67 was carried out by sandwich/fluorescence method. Brain sections were deparaffinized, demasked in citrate buffer pH 6.0 with 0.5% Tween-20 at 100 °C and blocked in phosphate-buffered saline with 0.1% bovine serum albumin at room temperature before exposure to antibodies (primary: Rabbit Polyclonal Anti-Ki67 Antibody, ab15580, Abcam, UK; secondary: Goat Anti-Rabbit IgG H&L Alexa Fluor 488 ab150077, Abcam). The nuclei were counterstained with DAPI (Sigma-Aldrich, USA). Digital images of peripheral, necrosis- and hemorrhage-free tumor regions (three fields of view at 200 × magnification) were captured using Zeiss Axioplan 2 fluorescence microscope in two channels with 488 nm and 358 nm excitation. The channels were merged in ImageJ software; Ki-67 index was determined as a percentage of Ki-67 positive cells among total tumor cell counts.
9
Immunofluorescence Localization of Mitochondrial and Metabolic Enzymes
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Cells were seeded at a density of 5–6 × 104 cells onto coverslips. After an attachment period, the cells were fixed with 4% paraformaldehyde for 20 min, washed three times with PBS (10 min), and permeabilized with 0.3% Triton X-100 for 15–20 min. After three washing steps, the cells were blocked for 60 min with 1% BSA and stained with the primary antibodies diluted 1:200 in 1% BSA anti-CPS1 [EPR7493-3] (Abcam, Cambridge, UK) and anti-COX IV (#11967S) (Cell Signaling Technology, Danvers, MA, USA) at 4 °C overnight. The cells were then washed three times with PBS and incubated with secondary antibodies goat anti-rabbit IgG H&L Alexa Fluor®488 (ab150077) and goat anti-mouse IgG H&L F(ab)2 Alexa Fluor®594 (#8890S) (Abcam, Cambridge, UK) for 2 h in the dark, at room temperature. Finally, cells were washed again with PBS. Next, cells were transferred onto a droplet of Vectashield® Mounting Medium with DAPI (Vector Laboratories, Burlingame, CA, USA) and sealed. Samples were analyzed using a Leica TCS SP5 confocal microscope (Bannockburn, IL, USA).
10
Quantification of CPS1 Expression by Flow Cytometry
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The cells (approximately 2.5 × 106 cells per sample) were seeded into Petri dishes. After an attachment period, cells were harvested into PBS and sedimented by low-speed centrifugation (500 g, 10 min, 4 °C). Next, cells were fixed by 3–4% formaldehyde at 37 °C for 10 min, centrifuged, washed with PBS, and permeabilized using 90% methanol on ice for 30 min. Subsequently, cells were washed with PBS. Nonspecific reactions were prevented by incubation in 3% BSA for 60 min. Cells were incubated with primary antibody anti-CPS1 [EPR7493-3] (Abcam, Cambridge, UK) diluted 1:80 in 3% BSA for 60 min. Then cells were washed with PBS and incubated 60 min with Goat anti-rabbit IgG H&L Alexa Fluor®488 (ab150077) (Abcam, Cambridge, UK) diluted 1:200 in 3% BSA. Finally, cells were washed with PBS, and the intensity of cell fluorescence was measured using a FACS Calibur cytometer (Becton Dickinson, San Jose, CA, USA).
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