Mouse control IgG (Santa Cruz Biotechnology, sc-2025) and rabbit control IgG (Millipore, 12–370), HRP-conjugated goat-anti mouse or rabbit IgG (Thermo Scientific, PA1-86717 and SA1-9510)(1:3,000), mouse anti-GFP (Sungene Biotech, KM8009)(1:1,000), mouse anti-FLAG (KM8002)(1:2,000), mouse anti-β-Actin (KM9001)(1:1,000), mouse anti-HA (COVANCE, MMS-101 R)(1:2,000), anti-pIκBα (9246L)(1:1,000), anti-Ubiquitin (sc-8017)(1:1,000), anti-IRF3 (sc-9082)(1:500), anti-IκBα (sc-371)(1:500), anti-p-IRF3 (4947 S)(1:1,000), anti-USP13 (abcam, GR56969-12)(1:500), anti-TBK1 (GR96328-11)(1:1,000) and anti-STING (13647 S)(1:1,000) were purchased from the indicated manufactures. Poly(I:C), ISD45, DNA90 and HSV120 were previously described36 (link)37 (link)38 (link)39 (link). ISD45: 5′-TACAGATCTACTAGTGATCTATGACTGATCTGTACATGATCTACA-3′; DNA90: 5′-TACAGATCTACTAGTGATCTATGACTGATCTGTACATGATCTACATACAGATCTACTAGTGATCTATGACTGATCTGTACATGATCTACA-3′; HSV120: 5′-AGACGGTATATTTTTGCGTTATCACTGTCCCGGATTGGACACGGTCTTGTGGGATAGGCATGCCCAGAAGGCATATTGGGTTAACCCCTTTTTATTTGTGGCGGGTTTTTTGGAGGACTT-3′.
Mouse anti ha
Manufactured by Fortrea
Sourced in United States, Germany
Mouse anti-HA is a monoclonal antibody that specifically binds to the hemagglutinin (HA) epitope tag. The HA tag is a commonly used protein tag for the identification and purification of recombinant proteins expressed in various systems.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti flag, by Merck Group (11 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (9 mentions)
Mouse anti β actin, by Merck Group (8 mentions)
Rabbit anti gfp, by Thermo Fisher Scientific (7 mentions)
Rabbit anti actin, by Merck Group (5 mentions)
Rabbit anti ha, by Cell Signaling Technology (5 mentions)
Cy3 conjugated goat anti mouse, by Jackson ImmunoResearch (4 mentions)
Sp5 confocal microscope, by Adobe (4 mentions)
Fitc conjugated goat anti hrp, by Jackson ImmunoResearch (4 mentions)
Mouse anti α tubulin, by Merck Group (4 mentions)
Chicken anti map2, by Abcam (4 mentions)
Rat anti ha, by Roche (4 mentions)
Alexa 488 conjugated goat anti rabbit, by Jackson ImmunoResearch (3 mentions)
Mouse anti fasciclin 2, by Developmental Studies Hybridoma Bank (3 mentions)
Mouse anti gfp, by Roche (3 mentions)
Mouse or rabbit fluorescent secondary antibody, by Jackson ImmunoResearch (3 mentions)
Rabbit anti cbp antibody, by Santa Cruz Biotechnology (3 mentions)
Anti flag m2, by Merck Group (3 mentions)
Mms 101p 500, by Jackson ImmunoResearch (3 mentions)
Mouse anti actin, by Merck Group (3 mentions)
96 protocols using mouse anti ha
1
Western Blotting Antibody Reagents
2
Detergent Lysis and Spreading of C. elegans Germ Cell Nuclei
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The protocol used for detergent lysis and spreading of C. elegans germ cell nuclei (S2 –S4 Figs, Figs 5 and 6 ) is an adapted version of the protocol used in [55 (link)] for S. cerevisiae. The gonads of 20–100 adult worms were dissected in 5μl dissection solution (see note A) on a 22x40mm coverslip (thickness #1.5, as required for the OMX microscope). 50μl of spreading solution (see note B) was added and gonads were immediately distributed over the whole coverslip using a glass rod or a pipette tip. Coverslips were left to dry overnight at room temperature or for two hours at 37°C, washed for 20 minutes in methanol at -20°C and rehydrated by washing 3 times for 5 minutes in PBS-T. A 20 minute blocking in 1% w/v BSA in PBS-T at room temperature was followed by overnight incubation with primary antibodies at 4°C (antibodies diluted in: 1% w/v BSA in PBS-T supplied with 0.05% w/v Sodium azide). Coverslips were washed 3 times for 5 minutes in PBS-T before secondary antibody incubation for 2 hours at room temperature. After PBS-T washes, the nuclei were immersed in Vectashield and the coverslip was mounted on a slide and sealed with nail polish. The following primary antibodies were used: chicken anti-HTP-3 (1:500 [46 (link)]), rabbit anti-GFP (1:750 [12 (link)]), guinea-pig anti-SYP-1 (1:200 [22 (link)]), rabbit anti-MSH-5 (1:10000 (SDIX)) and mouse anti-HA (Covance)
3
Characterizing Hepatitis C Virus Replication
4
Drosophila Embryo Antibody Staining
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Drosophila embryo collection, fixation, and antibody staining were carried out as previously described [15 ]. The following antibodies were used: FITC-conjugated goat anti-HRP (Jackson ImmunoResearch #123-095-021, 1:100), Alexa 647-conjugated goat anti-HRP (Jackson #123-605-021, 1:100), mouse anti-Fasciclin II (Developmental Studies Hybridoma Bank [DSHB] #1D4, 1:100), mouse anti-βgal (DSHB #40-1a, 1:150), mouse anti-Robo3 (DSHB #15H2, 1:100), mouse anti-HA (Covance #MMS-101P-500, 1:1000), rabbit anti-c-Myc (Sigma-Aldrich #C3956, 1:500), Cy3-conjugated goat anti-mouse (Jackson #115-165-003, 1:1000), Alexa 488-conjugated goat anti-rabbit (Jackson #111-545-003, 1:500). Embryos were genotyped using balancer chromosomes carrying lacZ markers. Ventral nerve cords from embryos of the desired genotype and developmental stage were dissected and mounted in 70% glycerol/PBS. Fluorescent confocal stacks were collected using a Leica SP5 confocal microscope and processed by Fiji/ImageJ [16 (link)] and Adobe Photoshop software.
5
Immunofluorescence Antibody Staining
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Primary antibodies used for immunofluorescence studies were as follows: mouse monoclonal anti-SipA (1:50), mouse monoclonal anti-SipB (1:50), mouse anti-FLAG M2 (Sigma-Aldrich, 1:1000), rabbit anti-GFP (Thermofisher, 1:50), mouse anti-HA (Covance, Clone 16B12, 1:400), goat anti-Salmonella CSA1 (KPL, 1:300) and rabbit anti-LAMP1 (Difco, 1:500). Alexa Fluor conjugated secondary antibodies were purchased from Life Technologies, diluted 1:1 in sterile glycerol (Sigma-Aldrich) and used at 1:400 dilution. Antibody dilutions specific to certain assays are listed with the appropriate method.
6
Quantitative Western Blotting of HA and ERp72
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HEK cells were transfected via jetPrime (Polyplus) by following the manufacture’s instructions. Cells were harvested in cold DPBS and lysed in ice-cold lysis buffer containing 25 mM Tris (pH 7.5), 2 mM MgCl2, 600 mM NaCl, 2 mM EDTA, 0.5% NP-40, 1X protease and phosphatase cocktail inhibitors (Sigma) and Benzonase (0.25U/μL of lysis buffer; Novagen). Aliquots of the proteins were separated by SDS-PAGE and then transferred to a polyvinylidene difluoride (PVDF) membrane (Amersham). After transfer, the membrane was washed 3X in Tris Buffer Saline (10 mM Tris-HCl pH 7.4, 150 mM NaCl) with 0.1% of Tween 20 (T-TBS), blocked for 1 h at room temperature in Odyssey Blocking Buffer (TBS, LI-COR), followed by 4 °C overnight incubation with the appropriate primary antibody in the above buffer. The following day, the membrane was washed 3X in T-TBS, incubated at room temperature for 1 h with IRDye secondary antibodies (LI-COR) at 1:10,000 dilution in Odyssey Blocking Buffer (TBS), followed by 3X T-TBS washes. Visualization was performed by quantitative fluorescence using an Odyssey CLx imager (LI-COR). Signal intensity was quantified using Image Studio software (LI-COR). Primary antibodies used for western-blotting are mouse anti-HA (1:2000, Covance), rabbit anti-ERp72 (1:1000, Cell Signaling Technologies).
7
Immunofluorescence Staining with Diverse Antibodies
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The following primary antibodies were used: chicken anti-HTP-3 (1:1000, (46 (link))), chicken anti-GFP (1:250, (Abcam)), mouse anti-GFP (1:500, (Roche)), rabbit anti-GFP (1:500, (47 (link))), mouse anti-HA (1:1000, (Clone 16B12; Covance)), rabbit anti-MSH-5 (1:10 000, (SDIX)), rabbit anti-RAD-51 (1:500, (48 (link))), rat anti-RAD-51 (1:200, (49 (link))), guinea pig anti-SUN-1 pS24 (1:700, (50 (link))), guinea pig anti-SUN-1 pS8(1:1000, (50 (link))), guinea pig anti-SYP-1 (1:200, (51 (link))). Secondary antibodies used were Alexa Fluor 405 (1:100), 488 (1:400), 555 (1:400), and 647 (1:200)-conjugated goat antibodies raised against the appropriate species (Life Technologies).
8
Fluorescent Staining of Drosophila Embryos
9
Co-immunoprecipitation of Harmonin and GSH
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Cells were lysed in 125 µl RIPA buffer for 20 minutes at 4°C. Proteins were extracted as previously
described (Blanco-Sánchez et al., 2014 (link)). After centrifugation at 5000 rpm for 5
minutes, the supernatant was incubated overnight at 4°C with either mouse anti-GSH (Virogen), rabbit anti-Ush1c (Novus
Biologicals) or rabbit anti-GFP (Torrey Pines Biolabs). Proteins were precipitated with protein A-Sepharose beads, washed in
lysis buffer and eluted in lysis buffer and non-reducing sample buffer (1:1). The eluted proteins were separated by
SDS-polyacrylamide gel electrophoresis and transferred to PVDF membranes. Detection was performed using rabbit anti-Harmonin
(Ush1c, Novus Biologicals, 1:600), mouse anti-HA (Covance, 1:500), rabbit anti-GFP (Torrey Pines Biolabs, 1:250), and mouse
anti-GSH (Virogen, 1:500) antibodies and the Odyssey Western Blotting Kit I LT (LI-COR) following the manufacturer
recommendations.
described (Blanco-Sánchez et al., 2014 (link)). After centrifugation at 5000 rpm for 5
minutes, the supernatant was incubated overnight at 4°C with either mouse anti-GSH (Virogen), rabbit anti-Ush1c (Novus
Biologicals) or rabbit anti-GFP (Torrey Pines Biolabs). Proteins were precipitated with protein A-Sepharose beads, washed in
lysis buffer and eluted in lysis buffer and non-reducing sample buffer (1:1). The eluted proteins were separated by
SDS-polyacrylamide gel electrophoresis and transferred to PVDF membranes. Detection was performed using rabbit anti-Harmonin
(Ush1c, Novus Biologicals, 1:600), mouse anti-HA (Covance, 1:500), rabbit anti-GFP (Torrey Pines Biolabs, 1:250), and mouse
anti-GSH (Virogen, 1:500) antibodies and the Odyssey Western Blotting Kit I LT (LI-COR) following the manufacturer
recommendations.
10
Immunohistochemical Analysis of Tret1-1
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Wandering third instar (L3) larval or adult brains were dissected and stained following standard protocols. Samples were imaged using a Zeiss LSM 880 (Zeiss, Oberkochen, Germany). The following antibodies were used: guinea pig anti-Tret1-1 PA (1:50, Volkenhoff et al., 2015 (link)), mouse anti-NC120 (1:2 Hybridoma), rabbit anti-laminin gamma (1:1,000 Abcam A47651), Chicken anti-GFP (1:500, Aves Labs), mouse anti-HA (1:1,000 Covance). Tret1-1 fluorescence was determined by comparing the mean gray values of Tret1-1 staining of null mutants or knockdown animals to the respective control. N is the number of independent experiments; n is the total number of animals analyzed.
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