The largest database of trusted experimental protocols
Sourced in United States, Germany, France, United Kingdom

The A11029 is a laboratory equipment product from Thermo Fisher Scientific. It is designed to perform a specific core function within a laboratory setting. However, without access to the technical specifications or detailed product information, I cannot provide a comprehensive and unbiased description of the product's capabilities or intended use. Therefore, the detailed description is not available.

Automatically generated - may contain errors

255 protocols using a11029

1

Immunohistochemical Analysis of Brain Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols
At 16 weeks post-implantation, animals were euthanized via transcardial perfusion and brains were extracted and prepared for sectioning as previously described64 ,65 (link).
For each analyzed marker, a minimum of 16 tissue sections from a minimum of four animals was used for statistical comparison. Immunohistochemical labeling of neuronal nuclei (NeuN), blood brain barrier stability (IgG) and astrocytes (GFAP) was performed using previously established methods47 (link),62 (link),65 (link). The following primary and secondary antibodies were used: Primary: Rabbit anti-glial fibrillary acidic protein (GFAP) (1:500, Z0334, Daco), mouse anti-neuronal nuclei (NeuN) (1:250, MAB377, Millipore), and rabbit anti-immunoglobulin G (IgG) (1:100, 618501, Bio-Rad). Secondary: Anti-mouse Alexa Fluor 488 (1:1000, A11029, LifeTechnologies) and anti-rabbit Alexa Fluor 594 (1:1000, A11037, LifeTechnologies).
+ Open protocol
+ Expand
2

Antibody Characterization and Procurement

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibody information can be found in Supplementary Table S1. Antibodies against CD68 (FA-11), Rab27A and V5 were purchased from Hycult Biotechnology (HM1070), Proteintech (17817-1-AP) and Life Technologies (R96025), respectively. Antibodies to GM130 (618022) and LAMP1 (553792) were purchased from BD Biosciences Pharmingen. Antibodies against α-tubulin (DM1A, T9026), β-actin (AC-15, A5441) and FLAG (F7425) were from SIGMA. Anti-Rab7 (D95F2, 9367) and anti-Rab11 (D4F5, 5589) were obtained from Cell Signaling. Antibodies against cathepsin K (E-7, sc-48353) and GFP (B-2, sc-9996) were from Santa Cruz. Antibodies against a1, a2 and a3 were generated as described previously30 ,61 (link),69 (link). Alexa-conjugated secondary antibodies (A11034, A11029, A11081 and A21236) and colloidal gold-conjugated ones (EMGAT10 and EMGMHL5) were from Life Technologies and BBI solutions, respectively. HRP-conjugated antibodies to rabbit IgG (NA934VS), mouse IgG (NA931VS), chicken IgY (12–341) and native primary antibodies (21230) were purchased from GE healthcare (anti-rabbit IgG and mouse IgG), Millipore and Thermo Scientific.
+ Open protocol
+ Expand
3

Immunofluorescence Staining of Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells of 0.2 million were seeded in chamber (154526, Thermo Scientific), and cultured for two days. Cells were fixed with cold methanol, membrane perforated with 0.15% Triton X-100 (TB0198, Sangon Biotech), blocked with animal non-immune serum (SP KIT-B, Maxvision), and incubated with primary antibodies overnight at 4℃, and then incubated with secondary antibodies for one hour. Cell nucleus was stained with DAPI (P36931, Life Technologies). Images were captured with confocal microscope (LSM 710, Zeiss) with 100x oil objective lens. The following antibodies were used for immune fluorescence: β-tubulin (1:100, HC101, TransGen Biotech), goat anti-mouse IgG secondary antibody Alexa Fluor 488 (1:200, A-11029, Life Technologies).
+ Open protocol
+ Expand
4

Immunofluorescence Protocol for Cellular Phenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols
To confirm the phenotype, zonula occludens-1 (ZO-1), cytokeratin-18 (CK18), and retinal pigment epithelium-specific 65 kDa protein (RPE65) immunofluorescence were used. The cells were fixed in 4% paraformaldehyde (PFA) (in phosphate buffer, PB) for 5 min at 4 °C. They were then transferred to 1% phosphate buffer saline (PBS) for washing purposes and incubated with a blocking buffer (1% bovine serum albumin, BSA, 0.5% Triton X-100, 0.2% sodium azide, and 1% FBS) for 1 h at 4 °C. The cells were incubated with an Alexa Fluor 594 mouse monoclonal antibody anti-ZO-1 (1:100, 339194, Invitrogen-Life Technologies, Gaithersburg, MD, USA), mouse monoclonal anti-CK18 (1:250, M7010, DAKO, Glostrup, Denmark), mouse monoclonal antibody to precursor and active MMP13 (1:250, MAB3321, Merck, Darmstadt, Germany), and rabbit polyclonal anti-RPE65 (1:250, NB100-355SS, Novus biologicals) at 4 °C overnight. After three 10 min washes, they were incubated with secondary fluorescent antibodies goat anti-mouse (1:250, A11029, Life technologies, Gaithersburg, MD, USA) and donkey anti-rabbit 594 (1:250, R37119, Invitrogen, Carlsbad, CA, USA) for 1 h in the dark. All antibody solutions were based on a blocking buffer. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Images were obtained using a laser scanning confocal microscope (LSM800, Zeiss, Oberkochen, Germany).
+ Open protocol
+ Expand
5

Phenotype Verification of ARPE-19 and HREC Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
To verify that ARPE-19 and HREC cells preserved their phenotype, we performed retinoid isomerohydrolase RPE65 (RPE65) (1:100, 78036, Abcam, Cambridge, MA, USA) and caveolin (1:250, 3238S, Cell Signaling, Danvers, MA, USA) staining by immunofluorescence. Briefly, 100,000 ARPE-19 and 50,000 HREC cells were seeded on a 10 mm dish (Menzel-Glaser, Waltham, MA, USA). Cold methanol was used for cellular fixing. Afterward, cells were washed with 1% phosphate buffer saline (PBS) and then incubated with blocking buffer containing 1% bovine serum albumin (BSA), 0.5% Triton X-100, 0.2% sodium azide, and 1% fetal bovine serum (FBS) for 1 h at 4 °C. Cells were incubated with the primary antibodies, diluted in blocking buffer at 4 °C for 24 h, and washed once more with PBS and then incubated with the secondary fluorescent antibodies goat anti-mouse 488 (1:250, A11029, Life technologies, Gaithersburg, MD, USA) and donkey anti-rabbit 488 (1:250, A21206, Invitrogen, Carlsbad, CA, USA) for RPE65 marker diluted in blocking buffer during 1 h in the dark. Nuclei were labelled with 4′,6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich, St. Louis, MO, USA). The morphology of cells was observed under an inverted phase-contrast microscope (Olympus CKX41, Tokyo, Japan) and photographed by a digital camera, and fluorescent images were obtained using a confocal microscope (LSM800, Zeiss, Oberkochen, Germany).
+ Open protocol
+ Expand
6

Immunofluorescence Microscopy of V5-tagged Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were prepared for immunofluorescence microscopy according to reference 14. The V5 epitope was detected with mouse anti-V5 monoclonal antibody SV5-Pk1 (1:750; AB27671; Abcam). The primary antibody was detected with an Alexa Fluor 488-conjugated goat anti-mouse polyclonal antibody (1:800; A11029; Life Technologies). Tubulin was labeled with a mouse anti-tubulin TAT1 monoclonal antibody (1:150) (41 (link)). The primary antibodies were detected with an Alexa Fluor 488-conjugated goat anti-mouse polyclonal antibody (1:250; A-11001; Life Technologies) or an Alexa Fluor 594-conjugated goat anti-rabbit antibody (1:250; catalog no. A-11037; Life Technologies). The cells were viewed with a Zeiss Axioplan 2 fluorescence microscope or a Zeiss 510 laser scanning confocal microscope. The images were processed with the Zen 2011 v7.0.0.285 software (Carl Zeiss GmbH) or the BioImageXD v1.0 RC3 software (42 (link)).
+ Open protocol
+ Expand
7

Immunofluorescence Staining of m-SKPs

Check if the same lab product or an alternative is used in the 5 most similar protocols
m-SKPs were snap frozen in optimal cutting temperature (OCT) compound (Fisher- LAMB/OCT) and sectioned at 8μm. Sections were either fixed in 4% paraformaldehyde (PFA) for 1 hour at room temperature and permeabilised with 0.1% Triton X-100, or fixed in ice-cold acetone for 30 minutes, and blocked with 3% bovine serum albumin (BSA) (Sigma-A2153). Primary antibodies were added in PBS containing 3% BSA and left at 4°C overnight. The following primary antibodies were used: Versican (1:10—DSHB—12C5), Fibronectin (1:100—Sigma—F3648), nestin (1:100—Millipore—MAB5326), βIII Tubulin (1:100—Sigma—T8660), S100β (1:100—Sigma—S2532) and α-SMA (1:100—Abcam—ab7817). Specimens were then washed in PBS before incubation with appropriate fluorophore conjugated secondary anti mouse IgG (Life Technologies—A11029) or anti rabbit IgG (Life Technologies—A11012) antibodies. Nuclei were stained with 4', 6-diamidino-2-phenylindole (DAPI) prior to mounting with Mowiol. Images were captured using Zeiss imager M1 fluorescent microscope with Velocity software (Improvision).
+ Open protocol
+ Expand
8

Immunofluorescence Analysis of CDH1 in BeWo Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Coverslips were cleaned by ethanol:chloric acid (99 : 1) wash. One coverslip was deposited into each well of a six-well plate and sterilized by UV treatment. BeWo cells were plated at a density of 50 000 cells per well. Cells after settling for 1 day were treated with DMSO or FSK for 48 h, washed in PBS twice at RT, and fixed in 2% PBS-PFA for 5 min at RT. The PFA was blocked by adding 0.125 M glycine for 5 min. After extensive PBS washing, cells were permeabilized with 0.1% PBS-Triton X-100 for 5 min. The slides were transferred to a humid chamber and, after 30 min blocking in PBS-BSA (10%) were incubated with primary antibody against CDH1 (ab1416, dilution 1/50, Abcam) in PBS-BSA (1%) overnight at 4 °C. Cells were washed and incubated with secondary antibody (A-11029, dilution 1/1000, Life Technologies) in PBS-BSA (1%) for 1 h at RT. After washing, DAPI was applied at a dilution of 1/10000. After extensive washing, the coverslips were mounted on slides with MOWIOL mounting medium. Fluorescent images were acquired using a Zeiss Axioplan 2 Imaging inverted fluorescence microscope in conjunction with a Zeiss Axiocam MRm monochromatic CCD camera or Zeiss Axiocam MRc color CCD camera and analyzed with Axiovision 4.8.2 software (Carl Zeiss AG, Oberkochen, Germany).
+ Open protocol
+ Expand
9

Subcellular Localization of P-FOXO and P-ERK

Check if the same lab product or an alternative is used in the 5 most similar protocols
Ae. aegypti Aag2 cells were seeded onto coverslips in 12-well plates at a confluency of approximately 1x106 cells/well. Cells were then treated for 24 hours with 1% DMSO, 1 μM DMAQ-B1, 10 μM AKT inhibitor VIII, or combined small molecule treatment supplemented in 2% FBS media as described [14 (link)]. Coverslips were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized in 0.1% Triton-X-100 for 30 minutes at room temperature and blocked in 1% BSA in TBS for 30 min at 37°C. Primary antibody labeling was completed with anti-P-FOXO (1:100) and anti-P-ERK (1:100) for 2 h at humified room temperature. Secondary antibody labeling was completed using anti-rabbit (Life Technologies A11034) or anti-mouse (Life Technologies A11029) Alexafluor 488 (1:300) by incubating membranes for 1 h at room temperature in the dark. Samples were stained with DAPI (1:100; Cell Signaling 4083), mounted onto coverslips using ProLong Diamond Antifade Mountant (Invitrogen P36961), and imaged using a Leica Sp8X confocal microscope. Localization percentages were determined using Adobe Illustrator 2021 by counting the total number of cells and evaluating if green-fluorescent signal was cytosolic, nuclear, or no signal in relation to DAPI-stained nuclei.
+ Open protocol
+ Expand
10

Immunofluorescence Staining of Human iPSC-Derived Hepatocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human iPSC derived hepatocytes were stained as previously described in Ulvestad et al. [15 (link)]. The primary antibodies used in this study were rabbit anti-α1-antitrypsin (1 : 200, A0012, DAKO), mouse anti-CK18 (1 : 100, M7010, DAKO), and rabbit anti-HNF4α (1 : 300, sc-8987, SantaCruz Biotechnology). The following secondary antibodies were used and purchased from Life Technologies: donkey anti-rabbit Alexa Fluor 488 IgG (1 : 1000, A21206), donkey anti-rabbit Alexa Fluor 594 IgG (1 : 1000, A21207), and goat-anti-mouse Alexa Fluor 488 (1 : 500, A11029).
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!