Deparaffinized tissue sections (5 μm) were permeabilized in PBS-T (1× PBS + 0.1% Triton X-100, three times for 5 min each), incubated with 5% BSA for 1 hour, and then stained for PAR (1:1000; catalog number 4335-MC-100, Invitrogen), HMGB1 (1:500; Ab18256, Abcam), 8-oxoG (1:200; Ab62623, Abcam), GFAP (1:500; Ab7779, Abcam), F4/80 (1:500; Ab6640, Abcam) or anti-pMLKL (1:200; Ab196436, Abcam) overnight at 4°C. After incubation with the secondary antibody conjugated with Alexa Fluor 488 or Alexa Fluor 568 (1:400; Invitrogen), nuclear counterstaining was performed using DAPI. Images were blindly analyzed using a Nikon Eclipse 6800 microscope, a Retiga Exi camera, and Velocity and ImageJ software.
Ab7779
Manufactured by Abcam
Sourced in United States
Ab7779 is a laboratory equipment product. It is designed for use in scientific research applications.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488, by Thermo Fisher Scientific (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Ab6640, by Abcam (1 mentions)
Alexa fluor 568, by Thermo Fisher Scientific (1 mentions)
Ab18256, by Abcam (1 mentions)
Ab62623, by Abcam (1 mentions)
Ab196436, by Abcam (1 mentions)
T2928, by Merck Group (1 mentions)
A11001, by Thermo Fisher Scientific (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
C5922, by Merck Group (1 mentions)
T5076, by Merck Group (1 mentions)
B2261, by Merck Group (1 mentions)
M1406, by Merck Group (1 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
Ab24567, by Abcam (1 mentions)
Ab7751, by Abcam (1 mentions)
Tcs sp8 confocal microscope, by Leica (1 mentions)
Ab11570, by Abcam (1 mentions)
Ab13847, by Abcam (1 mentions)
4 protocols using ab7779
1
Immunohistochemical Analysis of Oxidative Stress and Inflammation
2
Immunocytochemical Analysis of Neural Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Following flattening and adhesion, Hip-NSCs and their differentiated progeny were washed in PBS 2–3 times and incubated overnight at 4°C with the following primary antibodies: Rabbit anti-Nurr1 (1:1,000; N4663; Sigma-Aldrich), mouse anti-TH (1:8,000; T2928; Sigma-Aldrich), mouse anti-glial fibrillary acidic protein (GFAP; 1:4,000; SAB1405864; Sigma-Aldrich), rabbit anti-GFAP (1:3,000; ab7779; Abcam), mouse anti-β-Tubulin III (Tuj1; 1:3,000; T5076; Sigma-Aldrich), rabbit anti-Tuj1 (1:3,000; SAB4300623; Sigma-Aldrich) and mouse anti-2′, 3′-cyclic nucleotide 3′-phosphodiesterase (CNPas 1:500; C5922; Sigma-Aldrich). Cells were then incubated with the appropriate Alexa Fluor 488 or Alexa Fluor 555-labeled secondary antibodies (1:500; goat anti-mouse IgG; A11001; goat anti-rabbit IgG; A31629; Invitrogen; Thermo Fisher Scientific, Inc.). Nuclei were stained with Hoechst 33342 (1:500; B2261; Sigma-Aldrich). Coverslips were mounted onto slides, and observed under a fluorescence microscope.
3
Immunofluorescence Analysis of Spinal Cord Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At week 38 after operation, the spinal cord from T5-T11 was retrieved and fixed in 4% (v/v) formaldehyde for 48 h. The segments were then embedded in paraffin and cut into 10-μm thick sections (Leica Microsystems).
For immunofluorescence staining, the paraffin embedded sections were subjected to heat mediated antigen retrieval using citric acid, and the sections were then blocked using normal horse serum/T-PBS (1:20) for 15 min. Primary antibodies were applied to the sections and incubated overnight at 4 °C. The primary antibodies were against the following: GFAP (1:800, ab7779, Abcam), 5-HT (1:800, 20080, Immuostar), CS-56 (1: 500, ab11570, Abcam), Myelin Basic Protein (1: 500, ab24567, Abcam), Tuj-1/beta III Tubulin (1: 500, ab7751, Abcam), Map2 (1:500, M1406, Sigma), and SYN (1: 500, Ls-c174790, LSBio). Sections were then incubated with secondary antibody (Alexa Fluor 488, 1:400; Alexa Fluor 594, 1:800; Invitrogen). Cell nuclei were stained with DAPI (1:1000, sigma) and images were taken under the Leica TCS SP8 Confocal Microscope (Leica Microsystems, Germany). Image-Pro Plus software (Media Cybernetics LP, Maryland, USA) was used to quantify immunostaining positive signals by selecting at least 3 fields per sample at the lesion center.
For immunofluorescence staining, the paraffin embedded sections were subjected to heat mediated antigen retrieval using citric acid, and the sections were then blocked using normal horse serum/T-PBS (1:20) for 15 min. Primary antibodies were applied to the sections and incubated overnight at 4 °C. The primary antibodies were against the following: GFAP (1:800, ab7779, Abcam), 5-HT (1:800, 20080, Immuostar), CS-56 (1: 500, ab11570, Abcam), Myelin Basic Protein (1: 500, ab24567, Abcam), Tuj-1/beta III Tubulin (1: 500, ab7751, Abcam), Map2 (1:500, M1406, Sigma), and SYN (1: 500, Ls-c174790, LSBio). Sections were then incubated with secondary antibody (Alexa Fluor 488, 1:400; Alexa Fluor 594, 1:800; Invitrogen). Cell nuclei were stained with DAPI (1:1000, sigma) and images were taken under the Leica TCS SP8 Confocal Microscope (Leica Microsystems, Germany). Image-Pro Plus software (Media Cybernetics LP, Maryland, USA) was used to quantify immunostaining positive signals by selecting at least 3 fields per sample at the lesion center.
4
Antibody-Based Immunoblot and Immunofluorescence Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used for immunoblot analysis: rabbit anti-IKKβ [Y466] (ab32135, 1:2000; Abcam, Cambridge, United Kingdom), goat anti-Lcn2 (AF1857, 1:1000; R&D Systems, Minneapolis, MN, USA), rabbit anti-glial fibrillary acidic protein (GFAP; ab7779, 1:1000; Abcam), rabbit anti-Bax (2772, 1:1000; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-Bad (9292, 1:1000; Cell Signaling Technology), rabbit anti-NF-κB p65 (C20, sc-372, 1:1000; Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-phospho NF-κB p65 (Ser536, 3033, 1:1000; Cell Signaling Technology), rabbit anti-GAPDH (FL-335, (sc-25778, 1:2000; Santa Cruz Biotechnology), rabbit anti-β-tubulin (ab6046 1:10000; Abcam) and HRP-conjugated goat anti-rabbit or donkey anti-goat were obtained from Santa Cruz Biotechnology.
For immunofluorescence, the following primary antibodies were used: mouse anti-NeuN (MAB377, 1:300; Millipore-Sigma), rabbit anti-cleaved caspase 3 (ab13847, 1:400; Abcam), and mouse anti-tubulin-β3 (TUJ1) (MMS-435P, 1:500; BioLegend, San Diego, CA, USA). Corresponding Alexa Fluor-conjugated secondary antibodies were obtained from Thermo Fisher Scientific, and DAPI was purchased from Merck (Darmstadt, Germany).
For immunofluorescence, the following primary antibodies were used: mouse anti-NeuN (MAB377, 1:300; Millipore-Sigma), rabbit anti-cleaved caspase 3 (ab13847, 1:400; Abcam), and mouse anti-tubulin-β3 (TUJ1) (MMS-435P, 1:500; BioLegend, San Diego, CA, USA). Corresponding Alexa Fluor-conjugated secondary antibodies were obtained from Thermo Fisher Scientific, and DAPI was purchased from Merck (Darmstadt, Germany).
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!