A21428

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Japan

The A21428 is a laboratory instrument designed for analytical applications. It provides essential functionality for tasks involving sample analysis and measurement. The device specifications and core functions are presented in a factual and unbiased manner, without interpretation or extrapolation.

Automatically generated - may contain errors

Lab products found in correlation

123 protocols using a21428

Immunohistochemical Analysis of Neuronal Subtypes

Check if the same lab product or an alternative is used in the 5 most similar protocols

To mark PCs, brain sections were incubated with a rabbit anti-calbindin D28K antibody (1:1000, PA1-931, Invitrogen, Waltham, MA) at 4 °C for 24–48 h. After being washed in PBS, sections were incubated with a goat anti-rabbit Cy5 (1:500, A10523, Invitrogen) at room temperature for 3 h. After another rinse with PBS, sections were mounted on slides and cover-slipped with a mounting medium (H-1500, Vector Labs). To identify subtypes of c-Fos positive neurons following the social recognition test, sections were incubated with primary antibodies of guinea pig anti-c-Fos (1:1000, 226308, Synaptic Systems, Gottingen, Germany), rabbit anti-CaMKII (1:200, PA5-99558, Invitrogen) and mouse anti-GAD67 (1:500, MAB5406, Millipore, Burlington, MA) at 4 °C for 24–48 h. After being washed in PBS, sections were incubated with secondary antibodies of goat anti-guinea pig Alexa 488 (1:1000, A11073, Invitrogen), goat anti-rabbit Alexa 555 (1:1000, A21428, Invitrogen) and goat anti-mouse Alexa 647 (1:1000, A21236, Invitrogen) at room temperature for 3 h. After another rinse with PBS, sections were mounted on slides and cover-slipped with a mounting medium (H-1500, Vector Labs). Images were taken with a Zeiss LSM 710 confocal microscope and Zen 2.0 software (Oberkochen, Germany).

Chao O.Y., Pathak S.S., Zhang H., Augustine G.J., Christie J.M., Kikuchi C., Taniguchi H, & Yang Y.M. (2023). Social memory deficit caused by dysregulation of the cerebellar vermis. Nature Communications, 14, 6007.

+ Open protocol

+ Expand

Examining Synaptic Inputs in GnRH Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

To examine changes in kisspeptin and glutamatergic inputs to GnRH neurones across the oestrous cycle, POA and MBH tissue sections of luteal (n=4) and surge (n=4) animals were processed for triple-label immunodetection of kisspeptin, vGlut2 and GnRH, using the same protocol as above. Kisspeptin was visualised using TSA and CY5-steptavidin. GnRH neurones were detected using monoclonal mouse anti-GnRH (1:400; SMI41R, Sternberger Monoclonals, Inc., Princeton, NJ), and Alexa 488-goat anti-mouse IgG (1:100; A11001, Invitrogen). vGlut2 was visualised using Alexa555-goat anti-rabbit IgG (1:100; A21428, Invitrogen).

Merkley C.M., Coolen L.M., Goodman R.L, & Lehman M.N. (2015). Evidence for changes in numbers of synaptic inpcuts onto KNDy and GnRH neurones during the preovulatory LH surge in the ewe. Journal of neuroendocrinology, 27(7), 624-635.

+ Open protocol

+ Expand

Immunostaining of Murine ESCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Murine ESCs were plated onto glass coverslips in preparation for immunostaining. After 24 hours, cells were fixed with 4% paraformaldehyde in PBS, washed 3 times with PBS, and then permeabilized with 0.5% triton-X100 in PBS for 10 min at room temperature. Cells were then washed 3 times with PBS, blocked for 1 hour in 4% IgG-free BSA (Jackson ImmunoResearch 001-000-162) in PBS, and then stained over night with the indicated antibody in 4% IgG-free BSA at room temperature in a humidified chamber. Cells were then washed 3 times with PBS. Secondary antibodies were added to cells in 4% IgG-free BSA and incubated for 1 hour at room temperature. Cells were then washed twice in PBS. Cells were stained with Hoechst dye (Molecular Probes H3570) in PBS or water for 5 minutes, and then mounted in Vectashield mounting media (Vector Laboratories H-1000). Imaging was performed on an RPI spinning disk confocal at 100X magnification using the MetaMorph software v. 7.10.3.279 (Molecular Devices). Primary antibodies: anti-MAP2 (Invitrogen MA5-12823) and anti-MED1 (Abcam ab64965). Secondary antibodies: anti-mouse Alexa Fluor 568 (Invitrogen A11031) and antirabbit Alexa Fluor 555 (Invitrogen A21428).

Li C.H., Coffey E.L., Dall’Agnese A., Hannett N.M., Tang X., Henninger J.E., Platt J.M., Oksuz O., Zamudio A.V., Afeyan L.K., Schuijers J., Liu X.S., Markoulaki S., Lungjangwa T., LeRoy G., Svoboda D.S., Wogram E., Lee T.I., Jaenisch R, & Young R.A. (2020). MeCP2 links heterochromatin condensates and neurodevelopmental disease. Nature, 586(7829), 440-444.

+ Open protocol

+ Expand

Lectin Labeling Techniques for Vesicle Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

FITC-conjugated lectins from A. integrifolia (Jacalin; FL-1151) and D. stramonium (DSA; FL-1181) were purchased from Vector Laboratories (Burlingame, CA, USA) and FITC-conjugated lectins from A. caudatus (ACA; 21,510,290) were purchased from GlycoMatrix (Dublin, OH, USA). Colloidal gold-conjugated lectins Jacalin-15 nm gold (GP-6301–15), ACA-10 nm gold (GP-8201–10) and DSA-5 nm gold (GP-5701–5) were purchased from EY Laboratories (San Mateo, CA, USA). Lectin inhibitory molecule bovine submaxillary mucin for ACA, galactose for jacalin and chitin hydrolysate for DSA were also purchased from Vector Laboratories. Rabbit polyclonal anti-uroplakin (UP) primary antibody was a kind gift from Prof. Dr. Tung-Tien Sun, Department of Cell Biology, New York University Medical School (Wu et al. 1990 (link)). The following rabbit polyclonal antibodies were also used: anti-flotillin (ab41927, Abcam), anti-clathrin (610,499, BD Transductions), anti-caveolin (ab18199, Abcam) and anti-dynamin 2 (ab65556, Abcam). Goat anti-rabbit IgG AlexaFluor 488 (A11008, Invitrogen) and goat anti-rabbit IgG AlexaFluor 555 (A21428, Invitrogen) secondary antibodies were used. Vectashield mounting medium with DAPI, a nuclei marker, was obtained from Vector Laboratories.

Resnik N., Višnjar T., Smrkolj T., Kreft M.E., Romih R, & Zupančič D. (2023). Selective targeting of lectins and their macropinocytosis in urothelial tumours: translation from in vitro to ex vivo. Histochemistry and Cell Biology, 160(5), 435-452.

+ Open protocol

+ Expand

Immunofluorescence Imaging of FLAG-Tagged Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Flp-In T-REx HEK293 cells stably expressing 3XFLAG or BirA*-FLAG tagged proteins were seeded on poly-L-lysine coated cover slips (product #354085;BD Biosciences) at low density and grown in complete medium for 24 hours. Cells were fixed with 3.7% paraformaldehyde/PBS and permeabilized in 0.3% Triton X-100 in PBS. Mouse anti-FLAG M2 antibody (1:2000; F1804, Sigma-Aldrich), rabbit anti-FLAG antibody (1:1000; F7425, Sigma-Aldrich), mouse anti-centrin (1:1000, 04-1624, EMD Millipore), rabbit anti-pericentrin (1:2000, ab4448, Abcam) and streptavidin Alexa-Fluor 488 (1:2500; S32354; Invitrogen) were used to identify FLAG tagged proteins and biotinylated proteins, respectively. Proteins were visualized with goat anti-mouse or anti-rabbit coupled to Alexa-Fluor 488 or 555 antibodies (1:1,000; A11001, A11008, A21422, A21428; Invitrogen). DNA was detected with DAPI staining. Immunofluorescence was observed by confocal microscopy on a Nikon Eclipse C1si instrument.

Lambert J.P., Tucholska M., Go C., Knight J.D, & Gingras A.C. (2014). Proximity biotinylation and affinity purification are complementary approaches for the interactome mapping of chromatin-associated protein complexes. Journal of proteomics, 118, 81-94.

+ Open protocol

+ Expand

6mA Immunofluorescence Staining Protocol

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell fixation and immuofluoresence (IF) staining were performed as described previously (6 (link),39 (link),40 (link)). Cells were fixed during vegetative growth or at 10 h after mixing cells of two mating types, as indicated. For the 6mA staining, fixed cells were treated with RNase A (50 μg/ml or otherwise indicated) in phosphate-buffered saline (PBS) buffer for 2 h at 37°C. Non-treated cells were either incubated at 37°C or at room temperature (RT), as indicated. Cells were then incubated with 2N HCl for 20 min at RT and neutralized with Tris–HCl (100 mM, pH 8.5) for 10 min, prior to antibody incubation. The primary antibodies are α-6mA (Synaptic Systems, 202003, 1:2000) (5 (link),6 (link),10 (link)), α-HA (Cell Signaling, C29F4, 1:200), α-H3K9-acetylation (Abcam, ab10812, 1:500) and α-H3K14-acetylation (Abcam, ab52946, 1:500). The primary antibodies are incubated with cells at 4°C overnight (for α-6mA) or at RT for 2 h (for other antibodies). Cells were then incubated in the secondary antibody (Goat anti-Rabbit IgG (H+L), Invitrogen, A-21428, 1:4000) at RT for 1 h. Digital images were collected using a Leica DM2500 microscope with a Leica DFC450C camera.

Wang Y., Chen X., Sheng Y., Liu Y, & Gao S. (2017). N6-adenine DNA methylation is associated with the linker DNA of H2A.Z-containing well-positioned nucleosomes in Pol II-transcribed genes in Tetrahymena. Nucleic Acids Research, 45(20), 11594-11606.

+ Open protocol

+ Expand

Fluorescent Polystyrene Nanoparticles for FRET Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

The 40 nm-fluorescent carboxylated polystyrene (PS) nanoparticles (F-8789; ex: 660 nm/em: 680 nm) were purchased from Invitrogen (Carlsbad, CA). In our case, the fluorescence of the particles did not play a role in the FRET assay as discussed later; these particular particles were simply a suitable size and readily available. Goat-anti-Rabbit IgG-Alexa 555 (the donor molecule with excitation: 555 nm and emission : 565 nm; A21428) was purchased from Invitrogen. Rabbit IgG-Alexa 647 (the acceptor molecule with excitation: 650 nm and emission: 668 nm; SC-24647) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). 3-PBA hapten was synthesized. The detailed method was described previously [18 (link)]. Alexa 647 was conjugated to 3-PBA hapten by using a commercial protein labeling kit (A-20173, Invitrogen).

Han J.H., Sudheendra L, & Kennedy I.M. (2015). FRET-based homogeneous immunoassay on nanoparticle-based photonic crystal. Analytical and bioanalytical chemistry, 407(18), 5243-5247.

+ Open protocol

+ Expand

Immunofluorescence Analysis of Adipose Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fresh adipose tissues were isolated, post‐fixed and dehydrated with alcohol in increasing concentration: 70% for 24 h, 80% for 3 h, 90% for 4 h, 95% for 3 h and then absolute alcohol for 2 h. Adipose tissue slices with the thickness of 5 μm were cut by a microtome (Thermo Fisher Scientific), and blocked with 3% BSA, then, treated with primary antibodies TH (GeneTex, GTX634481, 1:200), YB‐1 (Cell signalling, 4202, 1:50). Subsequently, the adipose tissue slides were incubated with Alexa Fluor 488 conjugated anti‐Rabbit (Invitrogen, A21206, 1:2000) or Alexa Fluor 555 conjugated anti‐Rabbit (Invitrogen, A21428, 1:2000). All the fluorescence pictures were captured by a fluorescence microscope.

Zhou R., Huang Y., Feng X., Zhou R., Wang L., Xie G., Xiao Y, & Zhou H. (2023). Decreased YB‐1 expression denervates brown adipose tissue and contributes to age‐related metabolic dysfunction. Cell Proliferation, 57(1), e13520.

+ Open protocol

+ Expand

Immunofluorescent Staining of Organ Chips

Check if the same lab product or an alternative is used in the 5 most similar protocols

CRC Organ Chips were manually washed by flowing PBS through the endothelial and epithelial channels. The chips were fixed with 4% paraformaldehyde (Electron Microscopy Sciences, #15710), incubated for 15 minutes, and permeabilized with 1% saponin. Blocking buffer of 2% bovine serum albumin (BSA) and primary antibodies were incubated overnight at 4°C before a 2 hour incubation with secondary antibodies (1:100, Molecular Probes, Invitrogen, #A21428 and #A21235) diluted in blocking buffer. The primary antibodies used for these studies were anti-E-cadherin (1:25; Abcam, #ab1416), anti-VE-cadherin (1:25; Abcam, #ab33168), anti-vimentin (1:50; Abcam, #ab93547), anti-ZO-1 (1:100; Thermo-Fisher, #339194). DAPI (Sigma-Aldrich, D9542) was used to label all nuclei. Chips were imaged using the Perkin Elmer Operetta CLS High Content System. Quantitation of E-Cadherin+ and Vimentin+ (tumor cells identified by GFP) was performed using the Perkin Elmer Harmony software based on intensity thresholds.

Strelez C., Chilakala S., Ghaffarian K., Lau R., Spiller E., Ung N., Hixon D., Yoon A.Y., Sun R.X., Lenz H.J., Katz J.E, & Mumenthaler S.M. (2021). Human colorectal cancer-on-chip model to study the microenvironmental influence on early metastatic spread. iScience, 24(5), 102509.

+ Open protocol

+ Expand

Immunofluorescence Staining of BRD3 and TCOF1

Check if the same lab product or an alternative is used in the 5 most similar protocols

HeLa cells were seeded on coverslips at low density in Opti-MEM medium and then transfected with the appropriate siRNA using Lipofectamine RNAiMAX as per the manufacturer protocol, see STAR Methods and Table S1C for details. 48 hr later, cells were fixed with 3.7% paraformaldehyde/PBS and permeabilized in 0.3% Triton X-100 in PBS. Mouse anti-BRD3 antibody (1:100; ab50818, Abcam) and anti-TCOF1 (1:500; HPA038237; Sigma-Aldrich) were used to identify BRD3 and TCOF1, respectively. Proteins were visualized with goat anti-mouse or anti-rabbit coupled to Alexa Fluor 488 or 555 antibodies (1:1,000; A11001">A11001, A11008">A11008, A21422">A21422, A21428">A21428; Invitrogen). DNA was detected with DAPI staining. Immunofluorescence was observed by confocal microscopy on a Nikon Eclipse C1si instrument.

Lambert J.P., Picaud S., Fujisawa T., Hou H., Savitsky P., Uusküla-Reimand L., Gupta G.D., Abdouni H., Lin Z.Y., Tucholska M., Knight J.D., Gonzalez-Badillo B., St-Denis N., Newman J.A., Stucki M., Pelletier L., Bandeira N., Wilson M.D., Filippakopoulos P, & Gingras A.C. (2019). Interactome Rewiring Following Pharmacological Targeting of BET Bromodomains. Molecular Cell, 73(3), 621-638.e17.

+ 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?

Revolutionizing how scientists
search and build protocols!