The following primary antibodies were obtained from Developmental Studies Hybridoma Bank, DHSB, (Iowa City, IA): rat anti-DN-cadherin (DN-EX #8, adult, L3: 1:8, L1: 1:10), mouse anti-Brp (nc82,1:20), mouse anti-Neurotactin (BP106, 1: 6) mouse anti-Neuroglian (BP104, Adult: 1:12, L1: 1:14), mouse anti-repo (1:10), rabbit anti-βGal (1:3). We also used rabbit anti-HA (1:300, Cell Signaling Technologies), chicken anti-GFP (1:1000, Abcam) and rat anti-FLAG (1:300, Novus Biologicals). The following secondary antibodies were obtained from Jackson ImmunoResearch; Molecular Probes) and used: Alexa 568-conjugated anti-mouse (1:500), cy5-conjugated anti-mouse (1:300), cy3-conjugated anti-rat (1:300), Alexa 488- conjugated anti-rabbit (1:1000), cy5-conjugated anti-rat (1:300), cy3-conjugated anti-rabbit (1:160), and Alexa 488-conjugated anti-rabbit (1:300). We also obtained Alexa 488-conjugated anti-chicken (1:1000) from Thermo Fisher Scientific.
Alexa488 conjugated anti rabbit
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa488-conjugated anti-rabbit is a fluorescently labeled secondary antibody that specifically binds to rabbit primary antibodies. The Alexa488 fluorescent dye allows for the detection and visualization of target proteins in various applications such as immunofluorescence, flow cytometry, and Western blotting.
Automatically generated - may contain errors
Lab products found in correlation
Alexa 594 conjugated anti mouse antibodies, by Thermo Fisher Scientific (3 mentions)
Rabbit anti manf antibody, by Abcam (2 mentions)
Abc kit, by Vector Laboratories (2 mentions)
Lsm 510 meta, by Zeiss (2 mentions)
Ab4448, by Abcam (2 mentions)
Anti p53 sc 126, by Santa Cruz Biotechnology (2 mentions)
Anti gapdh a300 641a, by Fortis Life Sciences (2 mentions)
Anti γ h2ax 613402 clone 2f3, by BioLegend (2 mentions)
Anti 53bp1 h 300 sc 22760, by Santa Cruz Biotechnology (2 mentions)
Anti mpm 2 05 368mg, by Merck Group (2 mentions)
Anti h3s10p d2c8 3377, by Cell Signaling Technology (2 mentions)
Hpr linked anti mouse or anti rabbit nxa931 or na934v, by GE Healthcare (2 mentions)
Anti γ tubulin t6557, by Merck Group (2 mentions)
Ab80514, by Abcam (2 mentions)
Alexa 594 conjugated anti mouse, by Thermo Fisher Scientific (2 mentions)
Alexa488 conjugated anti goat, by Thermo Fisher Scientific (2 mentions)
Axioskop 2 plus, by Zeiss (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Lsm 510, by Zeiss (2 mentions)
Hrp conjugated anti rabbit and anti mouse secondary antibodies, by GE Healthcare (1 mentions)
16 protocols using alexa488 conjugated anti rabbit
1
Immunofluorescence Staining Protocol
2
Visualizing NS1-UAP56 Colocalization in Influenza
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A549 cells were infected with WT WSN or WSN-NS1-R38A-K41A mutant virus at a multiplicity of infection (MOI) of three. At the indicated time points post-infection, cells were fixed with 4% paraformaldehyde in PBS and permeabilized with 0.1% Triton X-100 in PBS. Mouse anti-UAP56 antibody (LS-C172345; LSBio) and rabbit anti-NS1 antibody (PA5-32243; Thermo Fisher) were used as primary antibodies. Alexa 488-conjugated anti-rabbit and Alexa 594-conjugated anti-mouse antibodies (Life Technologies) were used as secondary antibodies. Slides were mounted in mounting media with DAPI, and analyzed by using LSM510 META (Carl Zeiss). The immunofluorescence co-localization of NS1 and UAP56 was assessed by Pearson’s correlation coefficient of red- and green-pixels, calculated by using the FIJI “coloc2” function1 (Schindelin et al., 2012 (link)).
3
Immunohistochemical Analysis of MANF Expression
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Rabbit anti-MANF antibody was purchased from Abcam (Cambridge, MA). Recombinant MANF protein was expressed and purified as previously described [10] (link). Mouse anti-neuronal nuclei (NeuN) antibody was obtained from Millipore Corporate (Billerica, MA). Mouse anti-glial fibrillary acidic protein (GFAP) and calbindin antibodies were obtained from Sigma Chemical Co. (St. Louis, MO). Mouse anti-tyrosine hydroxylase antibody was purchased from BD biotechnology (San Diego, CA). Biotin-conjugated anti-mouse and anti-rabbit secondary antibodies and the ABC kit were obtained from Vector (Burlingame, CA). Alexa-488 conjugated anti-rabbit and Alexa-594 conjugated anti-mouse antibodies were obtained from Life Technologies (Grand Island, NY). Mouse anti-actin, HRP-conjugated anti-rabbit and anti-mouse secondary antibodies were purchased from GE Healthcare Life Sciences (Piscataway, NJ). Ketamine/xylazine was obtained from Butler Schein Animal Health (Dublin, OH). Other chemicals and reagents were purchased either from Sigma Chemical or Life Technologies.
4
Immunoblotting and Immunohistochemistry of ER Stress Markers
5
Immunofluorescent Labeling of Microglial Cells
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Microglial cells were plated on poly-L-lysine coated coverslips. After treatments, cells were then fixed with paraformaldehyde (4% in PBS) for 20 min at room temperature, and permeabilized for 5 min in PBS containing 0.3% Triton X100. Following 3 washings in PBS, the blocking step was realized with 3% BSA in PBS at room temperature for 30 min. The cells were then incubated overnight at 4°C with a rabbit anti-Iba1 (1:300, Biocare Medical, USA). After washing steps with PBS, cells were incubated with Alexa488-conjugated anti-rabbit (1:1000, Life Technologies, Belgium) at room temperature for one hour. The cells were washed and mounted with Dapi-Fluoromount G (SouthernBiotech, USA). Cells were observed under a LSM 510 META inverted confocal microscope (Carl Zeiss Micro Imaging, Göttingen, Germany) at a 40-fold magnification.
6
Immunofluorescence Assay for DNA Damage Response
7
Immunofluorescence Assay for DNA Damage Response
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Primary antibodies: anti-γ-H2AX (613402 clone 2F3, Biolegend); anti-TRF1 (Karlseder lab); anti-TRF2 (Karlseder lab); anti-53BP1 (H-300) (sc-22760, Santa Cruz); anti-LIG4 (ab80514, Abcam); anti-pericentrin (ab4448, Abcam); anti-MPM-2 (05-368MG, Millipore); anti-γ-tubulin (T6557, Sigma-Aldrich); anti-H3S10P (D2C8) (3377, Cell Signaling); anti-GAPDH (A300-641A, Bethyl); anti-p53 (sc-126, Santa Cruz).
Secondary antibodies: HPR-linked anti-mouse or anti-rabbit (NXA931 or NA934V; GE Healthcare); Alexa-488-conjugated anti-rabbit (Invitrogen); Alexa-594-conjugated anti-mouse (Invitrogen).
Secondary antibodies: HPR-linked anti-mouse or anti-rabbit (NXA931 or NA934V; GE Healthcare); Alexa-488-conjugated anti-rabbit (Invitrogen); Alexa-594-conjugated anti-mouse (Invitrogen).
8
Antibody-based analysis of autophagy and apoptosis
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Antibodies against the following proteins were used: ATG5 (#0262-100/ATG5-7C6) from Nanotools (Teningen, Baden-Württemberg, DE), ATG7 (#8558) ), Cleaved-Caspase3 (#9664), LAMP-1 (#9091) from Cell Signaling Technology (Danvers, MA, USA), PARP-1 (C-2-10) (#BML-SA249-0050) from Enzo Life Sciences (Farmingdale, NY, USA), Actin β (#NB600-501) from Novus Biologicals (Centennial, CO, USA), HRP conjugated rabbit (#111-035-003) and HRP conjugated mouse (#115-035-174) from Jackson ImmunoResearch (Ely, Cambridgeshire, UK), Alexa488 conjugated anti-rabbit (#A11008) from Invitrogen (Waltham, MA, USA). Hoechst 33258 (#14530), E64d (#E8640), Propidium iodide (#P4864), Chloroquine diphosphate salt (#C6628), QV-D-Oph (#SML0063), butylhydroxyanisole (BHA, #B1212000), N-acetyl-L-cysteine (NAC, #A8199) were purchased from Sigma-Aldrich (St. Louis, MO, USA).). Tetramethylrhodamine methyl-ester, TMRM (T-668) was purchased from Molecular Probes (Eugene, OR, USA). Fluoromount G (#00-4958-02) was purchased from Invitrogen. 20A was synthesized as previously described [42 (link),43 (link)]. Ferrostatin-1 (#341494) was purchased from Calbiochem (San Diego, CA, USA)), while LLOME (#16008) and Erastin (#17754) were purchased from Cayman Chemical (Ann Arbor, MI, USA).
9
Immunofluorescence Staining of Cortical Neurons
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Immunofluorescence staining was performed as described previously (Fanibunda et al., 2019 (link)). In brief, cortical neurons were fixed in 4% paraformaldehyde, followed by blocking in 10% horse serum and incubated with primary antibodies, rabbit anti-pCREB (1:1000; Cell Signaling Technology, MA, United States) or goat anti-5-HT2A receptor (1:500, Santa Cruz Biotechnologies, United States) along with the pan-neuronal marker, mouse anti-MAP2 (1:1000, Sigma-Aldrich, United States) overnight at 4°C. This was followed by incubation with secondary antibodies, Alexa 488 conjugated anti-goat (1:500; Molecular probes, CA, United States) or Alexa 488 conjugated anti-rabbit (1:500; Molecular Probes, CA, United States) or biotinylated horse anti-mouse (1:500, Roche Applied Science, Switzerland) with subsequent incubation with streptavidin-conjugated Alexa 568 (1:500, Molecular Probes, CA, United States) for 2 h. Following secondary antibody incubation and serial washes, cortical neurons were mounted in Vectashield (Vector Laboratories, CA, United States), and images were captured on the Zeiss LSM5 Exciter laser scanning microscope.
10
Immunofluorescence Analysis of STAT3 in HepG2 Cells
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HepG2 cells were grown on polylysine-coated coverslips for 24 h in the presence of Modified Eagle medium supplemented with 10% fetal bovine serum. Appropriate treatments were given to cells. Then cells were washed with 1XPBS, fixed with 4% paraformaldehyde and permeabilized with 0.5% Triton X-100. Anti-STAT3 (Abcam) antibody was used as primary antibody and fluorescently labeled anti-Rabbit Alexa-488 conjugated (Invitrogen) was used as secondary antibody. Hoechst and Alexa fluorescence were visualized using a Carl Zeiss confocal laser scanning microscope (Axioskop 2 Plus).
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