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Habp013

Manufactured by Merck Group
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HABP013 is a laboratory equipment product manufactured by Merck Group. It is designed for general laboratory use. The core function of HABP013 is to perform specific tasks related to laboratory procedures, but detailed information about its intended use is not available.

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Lab products found in correlation

Mab2b, by Synaptic Systems (2 mentions) Dylight conjugates, by Jackson ImmunoResearch (2 mentions) Vectashield mounting medium, by Vector Laboratories (2 mentions) Anti pkc, by Merck Group (2 mentions) Vectashield antifade mounting medium, by Vector Laboratories (2 mentions) Anti dsred, by Takara Bio (1 mentions) Anti ctbp2, by BD (1 mentions)

5 protocols using habp013

1

Whole-Mount Immunolabeling of Retinal Neurons

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Animals were euthanized with Isoflurane, decapitated, and enucleated. Retinas were dissected in cold oxygenated mouse artificial cerebrospinal fluid (mACSF, pH 7.4) containing (in mM): 119 NaCl, 2.5 KCl, 2.5 CaCl2, 1.3 MgCl2, 1 NaH2PO4, 11 glucose, and 20 HEPES. During retina dissections note was taken of retinal location and orientation (Wei et al., 2010 (link)). Whole-mount immunolabelings were performed on retinas that were flattened on a filter paper (Millipore, HABP013). Flat-mounted retinas were fixed for 15 mins in 4% paraformaldehyde (in mACSF). After rinses in phosphate buffer (PBS), the retinas were pre-incubated in blocking solution containing 5% donkey serum and 0.5% Triton X-100 (in PBS) followed by incubation with primary antibodies in blocking solution over 3–4 nights at 4°C. Primary antibodies used were directed against: GFP (1:1000, chicken, Abcam), GABAAα3 (1:3000, guinea pig, J.M. Fritschy), GABAAα1 (1:5000, guinea pig, J.M. Fritschy), GlyRα1 (1:500, mouse monoclonal mAb2b, Synaptic Systems) and gephyrin (1:1000, mouse monoclonal clone 3B11, Synaptic systems). Secondary antibody incubation was carried out overnight in PBS using anti-isotypic Alexa Fluor (1:1000, Invitrogen) or DyLight conjugates (1:1000, Jackson Immunoresearch). Retinas were subsequently mounted on slides using Vectashield mounting medium (Vector Labs).
Sawant A., Ebbinghaus B.N., Bleckert A., Gamlin C., Yu W.Q., Berson D., Rudolph U., Sinha R, & Hoon M. (2021). Organization and emergence of a mixed GABA-glycine retinal circuit that provides inhibition to mouse ON-sustained alpha retinal ganglion cells. Cell reports, 34(11), 108858.
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2

Immunostaining of Mouse Retina Tissue

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Retinas were isolated in cold oxygenated mouse artificial cerebrospinal fluid (mACSF, pH 7.4, 119 mM NaCl, 2.5 mM KCl, 2.5 mM CaCl2, 1.3 mM MgCl2, 1 mM NaH2PO4, 11 mM glucose, and 20 mM HEPES). Retinas were flattened onto filter paper (Millipore, HABP013) and fixed for 15 mins in 4% (wt/vol) paraformaldehyde prepared in mACSF. Retinas were rinsed in phosphate buffer (PBS) and then incubated in a blocking solution (5% donkey serum and 0.5% Triton X-100). The retinas were next incubated with primary antibody over 3 nights at 4°C. Primary antibodies used were anti-PKC (1:1000, mouse, Sigma; RRID:AB_477375), anti-Dsred (rabbit 1:1000, Clontech), anti-synaptotagmin 2 (1:1000, mouse, Znp-1 Zebrafish International Resource center; RRID:AB_10013783), anti-calbindin antibody (rabbit, 1:1000, Swant Inc; RRID:AB_10000340), anti-GABAAβ2/3 (mouse, 1:500 MilliporeSigma) anti-GABAAα1 receptor subunit (polyclonal guinea-pig, 1:5000, kindly provided by J.M. Fritschy), and anti-GABACρ receptor subunit (1:500, rabbit, kindly provided by R. Enz, H. Wassle, and S. Haverkamp). Retinas were thereafter incubated in secondary antibody solution using anti-isotypic Alexa Fluor (1:1000, Invitrogen) conjugates. Retinas were finally mounted on slides with Vectashield antifade mounting medium (Vector Labs; RRID:AB_2336789).
Nagy J., Ebbinghaus B., Hoon M, & Sinha R. (2021). GABAA presynaptic inhibition regulates the gain and kinetics of retinal output neurons. eLife, 10, e60994.
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3

Immunohistochemical Analysis of Mouse Retina

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Mice were deeply anesthetized with Isoflurane (5%), cervically dislocated or decapitated, and enucleated. Retinas were dissected in room temperature oxygenated mouse artificial cerebral spinal fluid (mACSF, pH 7.4) containing (in mM): 119 NaCl, 2.5 KCl, 2.5 CaCl2, 1.3 MgCl2, 1 NaH2PO4, 11 glucose, and 20 HEPES. Retinas were flat mounted on filter paper (Millipore, HABP013) then fixed for 15 minutes in 4% paraformaldehyde in mACSF. Retinas were rinsed with PBS, 3-10 minute washes, and incubated in blocking solution containing 5% normal donkey serum and 0.5% Triton X-100 overnight at 4°C. The retinas were then incubated with primary antibodies in blocking solution for 3 nights at 4°C. After rinsing in PBS, 3-10 minutes each, and the retinas were incubated in secondary antibodies in PBS solution overnight at 4°C. The tissues were rinsed with PBS prior to mounting on slides with Vectashield for confocal imaging.
Gamlin C.R., Zhang C., Dyer M.A, & Wong R.O. (2020). Distinct developmental mechanisms act independently to shape biased synaptic divergence from an inhibitory neuron. Current biology : CB, 30(7), 1258-1268.e2.
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4

Immunolabeling of Retinal Whole-Mounts

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Immunolabeling was performed on whole-mount retinas isolated in cold oxygenated mouse artificial cerebrospinal fluid (mACSF, pH 7.4) containing (in mM): 119 NaCl, 2.5 KCl, 2.5 CaCl2, 1.3 MgCl2, 1 NaH2PO4, 11 glucose, and 20 HEPES. Retinas were flattened on a filter paper (Millipore, HABP013), fixed for 15 mins in 4% paraformaldehyde prepared in mACSF, rinsed in phosphate buffer (PBS) and incubated with primary antibody in blocking solution containing 5% donkey serum and 0.5 Triton X-100 at 4°C for 3–4 days. Antibodies utilized were as follows: anti-PKC (1:1000, mouse, Sigma); anti-LRRTM4 (BC-262) (1:500, rabbit, Siddiqui et al., 2013 (link)); anti-VIAAT (1:1000, guinea pig, Synaptic Systems); anti-GABAAα1 (1:5000, guinea pig, J.M Fritschy); anti-GABACρ (1:500, rabbit, R. Enz, H. Wassle and S. Haverkamp); anti-CtBP2 (1:1000, mouse, BD Biosciences), anti-GABAAα3 (1:3000, guinea pig, J.M Fritschy). After incubation with primary antibodies, retinas were rinsed in PBS and incubated with anti-isotypic Alexa Fluor (1:1000, Invitrogen) or DyLight (1:1000, Jackson Immunoresearch) secondary antibodies overnight at 4°C. Thereafter retinas were rinsed in PBS and mounted on slides with Vectashield antifade mounting medium (Vector Labs).
Sinha R., Siddiqui T.J., Padmanabhan N., Wallin J., Zhang C., Karimi B., Rieke F., Craig A.M., Wong R.O, & Hoon M. (2020). LRRTM4: A novel regulator of presynaptic inhibition and ribbon synapse arrangements of retinal bipolar cells. Neuron, 105(6), 1007-1017.e5.
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5

Whole-Mount Immunolabeling of Retinal Neurons

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Animals were euthanized with Isoflurane, decapitated, and enucleated. Retinas were dissected in cold oxygenated mouse artificial cerebrospinal fluid (mACSF, pH 7.4) containing (in mM): 119 NaCl, 2.5 KCl, 2.5 CaCl2, 1.3 MgCl2, 1 NaH2PO4, 11 glucose, and 20 HEPES. During retina dissections note was taken of retinal location and orientation (Wei et al., 2010 (link)). Whole-mount immunolabelings were performed on retinas that were flattened on a filter paper (Millipore, HABP013). Flat-mounted retinas were fixed for 15 mins in 4% paraformaldehyde (in mACSF). After rinses in phosphate buffer (PBS), the retinas were pre-incubated in blocking solution containing 5% donkey serum and 0.5% Triton X-100 (in PBS) followed by incubation with primary antibodies in blocking solution over 3–4 nights at 4°C. Primary antibodies used were directed against: GFP (1:1000, chicken, Abcam), GABAAα3 (1:3000, guinea pig, J.M. Fritschy), GABAAα1 (1:5000, guinea pig, J.M. Fritschy), GlyRα1 (1:500, mouse monoclonal mAb2b, Synaptic Systems) and gephyrin (1:1000, mouse monoclonal clone 3B11, Synaptic systems). Secondary antibody incubation was carried out overnight in PBS using anti-isotypic Alexa Fluor (1:1000, Invitrogen) or DyLight conjugates (1:1000, Jackson Immunoresearch). Retinas were subsequently mounted on slides using Vectashield mounting medium (Vector Labs).
Sawant A., Ebbinghaus B.N., Bleckert A., Gamlin C., Yu W.Q., Berson D., Rudolph U., Sinha R, & Hoon M. (2021). Organization and emergence of a mixed GABA-glycine retinal circuit that provides inhibition to mouse ON-sustained alpha retinal ganglion cells. Cell reports, 34(11), 108858.
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