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Alexa 647 donkey anti mouse igg

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Alexa Fluor 647 donkey anti-mouse IgG is a secondary antibody conjugate used for the detection and visualization of mouse immunoglobulin G (IgG) in various immunoassays and imaging applications. The antibody is produced in donkeys and conjugated to the fluorescent dye Alexa Fluor 647, which has excitation and emission maxima of 650 nm and 665 nm, respectively.

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Lab products found in correlation

Lsm800 confocal microscope, by Zeiss (2 mentions) Prolong gold mounting medium, by Thermo Fisher Scientific (2 mentions) Mouse anti nestin, by Abcam (2 mentions) Chicken anti neurofilament m h, by Aves Labs (2 mentions) Alexa 488 donkey anti chicken igg, by Jackson ImmunoResearch (2 mentions) Alexa 405 donkey anti guinea pig igg, by Jackson ImmunoResearch (2 mentions) Rabbit anti rfp, by Rockland Immunochemicals (2 mentions) Guinea pig anti synapsin1 2, by Synaptic Systems (2 mentions) Alexa 594 donkey anti rabbit igg, by Jackson ImmunoResearch (2 mentions) Cy3 donkey anti mouse igg, by Jackson ImmunoResearch (1 mentions) Donkey anti mouse igg alexa 488, by Jackson ImmunoResearch (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Mowiol 4 88, by Merck Group (1 mentions) Phospho histone h3, by Abcam (1 mentions) Click it edu kit, by Thermo Fisher Scientific (1 mentions) Mef2c, by Santa Cruz Biotechnology (1 mentions) Lsm 700, by Zeiss (1 mentions) α tubulin, by Merck Group (1 mentions) Integrin β1, by Abcam (1 mentions) Goat anti mouse igg cy3, by Jackson ImmunoResearch (1 mentions)

Close spelling variants of this product

Alexa 647 (67 citations) Donkey anti-mouse Alexa 647 (11 citations) Alexa Fluor 647 donkey anti-mouse IgG (11 citations) Donkey anti-mouse 647 (7 citations) Alexa Fluor 647–conjugated donkey anti-mouse IgG (6 citations) Alexa Fluor 647 AffiniPure donkey anti-mouse IgG (6 citations) Alexa Fluor® 647 AffiniPure Donkey Anti-Mouse IgG (H+L) (3 citations)

4 protocols using alexa 647 donkey anti mouse igg

1

Immunohistochemistry and EdU Staining Protocol

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We collected 10 μm-thick cryosections in accordance with standard procedure for Immunohistochemistry and EdU staining. When combined with EdU detection, we carried out firstly EdU staining using Click-iT EdU kit (Invitrogen, C10340) in accordance with the manufacturer’s instructions, then followed by a standard immunohistochemistry protocol as previously described (Fei et al., 2017 (link)). In brief, after PBS wash and permeabilization with PBST, slides were blocked in 5% serum prepared in PBST, then incubated with primary antibodies overnight at 4°C, followed by sequential PBST washes and secondary antibody (with DAPI, Sigma-Aldrich, D9542) incubation, and finally mounted using Mowiol 4-88 (Sigma-Aldrich, 9002-89-5) mounting medium after several PBST washes. Fluorescence images were acquired with an Olympus IX83 microscope (Olympus, Tokyo, Japan), using ×20 objectives and analyzed using FIJI. The following antibodies were used in this study, MBP (Genetex, GTX761141), PRRX1 (Gerber et al., 2018 (link)), βIII-TUBULIN (R&D, MAB1195), SOX9 (Chemicon, Ab5535), PAX7 (DSHB, AB528428), MEF2C (Santa Cruz, sc-365862), phospho-Histone H3 (Abcam, 10543), MHC (DSHB, A4.1025). Alexa 488-donkey-anti-mouse IgG (Jackson, 711-547-003), Alexa 555-donkey-anti-rat IgG (Invitrogen, SA5-10027), Alexa 647-donkey-anti-mouse IgG (Jackson, 715-607-003), CY3-donkey-anti-mouse IgG (Jackson, 715-165-151).
Hu Y., Pan X., Shi Y., Qiu Y., Wang L., Murawala P., Liu Y., Xing W., Tanaka E.M, & Fei J.F. (2022). Muscles are barely required for the patterning and cell dynamics in axolotl limb regeneration. Frontiers in Genetics, 13, 1036641.
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2

Immunohistochemical Detection of Neural Markers

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For immunohistochemistry, fixed coverslips were incubated in blocking solution (3% normal donkey serum, 0.3% Triton X-100 in TBS) at room temperature for 1 hour. Primary antibodies, mouse anti-nestin (1:500, Abcam), guinea pig anti-synapsin1/2 (1:500; Synaptic Systems), rabbit anti-RFP (1:500; Rockland), or chicken anti-neurofilament (M+H; 1:1000; Aves Labs) were diluted in antibody diluent solution (1% normal donkey serum in 0.3% Triton X-100 in TBS) and incubated overnight at 4°C. Samples were then rinsed three times in TBS and incubated in secondary antibody solution (Alexa 488 donkey anti-chicken IgG; Alexa 594 donkey anti-rabbit IgG, Alexa 647 donkey anti-mouse IgG, and Alexa 405 donkey anti-guinea pig IgG all used at 1:500 (Jackson Immuno Research)) in antibody diluent solution at 4°C overnight. Coverslips were rinsed three times in TBS and mounted with ProLong Gold Mounting medium (Life Technologies). Images were collected using a 63x oil-immersion objective on a Zeiss LSM800 confocal microscope and processed with airyscan. Coverslips were used to quantify colocalization as described below.
Venkatesh H.S., Morishita W., Geraghty A.C., Silverbush D., Gillespie S.M., Arzt M., Tam L.T., Espenel C., Ponnuswami A., Ni L., Woo P.J., Taylor K.R., Agarwal A., Regev A., Brang D., Vogel H., Hervey-Jumper S., Bergles D.E., Suvà M.L., Malenka R.C, & Monje M. (2019). Electrical and synaptic integration of glioma into neural circuits. Nature, 573(7775), 539-545.
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3

Immunofluorescence Staining of 3D Cell Cultures

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Cells from three-dimensional cultures were fixed with 2% paraformaldehyde/PBS for 10 min, permeabilized in 0.5% Triton X-100/10% Goat serum/10% fish gelatin/PBS for 1 h and incubated overnight in primary antibodies. The primary antibodies were used at the following dilutions: Par6B 1/200 (Santa Cruz #sc-67393), aPKCι 1/100 (BD Transduction #610175), Par3 1/200 (Millipore #07-330), CD13 1/100 (Abcam #ab108382), CD13 1/50 (Sigma #HPA004625), E-cadherin 1/200 (Cell Signaling #3195 S), ZO-1 1/100 (Cell Signaling #8193), Ezrin 1/200 (Cell Signaling #3145), Pals1 1/100 (proteintech group #17710-1-AP), β1-integrin 1/200 (Abcam #ab30394), V5 1/200 (Thermo Fisher Scientific # R960-25), laminin 1/200 (Abcam #ab11575), α-Tubulin 1/200 (Sigma #T9026), Phalloidin 1/100 (Invitrogen #A34055), and endoplasmic reticulum (ER) marker-Calreticulin 1/200 (Abcam #ab92516). The secondary antibodies conjugated to Alexa488-Donkey anti-Rabbit IgG (Jackson ImmunoResearch Laboratories #711-545-152), Goat Anti-Mouse IgG (Cy3) (Jackson ImmunoResearch Laboratories #115-165-166) and Alexa647-Donkey Anti-Mouse IgG (Jackson ImmunoResearch Laboratories #715-605-151) were used at 1:750. DNA was detected with Hoechst dye 33258. Confocal imaging was performed using LSM700 from Zeiss with 20×/0.8NA or 40×/1.4NA objective lenses and processed using FIJI/ImageJ software.
Wang L.T., Rajah A., Brown C.M, & McCaffrey L. (2021). CD13 orients the apical-basal polarity axis necessary for lumen formation. Nature Communications, 12, 4697.
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4

Immunohistochemical Detection of Neural Markers

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For immunohistochemistry, fixed coverslips were incubated in blocking solution (3% normal donkey serum, 0.3% Triton X-100 in TBS) at room temperature for 1 hour. Primary antibodies, mouse anti-nestin (1:500, Abcam), guinea pig anti-synapsin1/2 (1:500; Synaptic Systems), rabbit anti-RFP (1:500; Rockland), or chicken anti-neurofilament (M+H; 1:1000; Aves Labs) were diluted in antibody diluent solution (1% normal donkey serum in 0.3% Triton X-100 in TBS) and incubated overnight at 4°C. Samples were then rinsed three times in TBS and incubated in secondary antibody solution (Alexa 488 donkey anti-chicken IgG; Alexa 594 donkey anti-rabbit IgG, Alexa 647 donkey anti-mouse IgG, and Alexa 405 donkey anti-guinea pig IgG all used at 1:500 (Jackson Immuno Research)) in antibody diluent solution at 4°C overnight. Coverslips were rinsed three times in TBS and mounted with ProLong Gold Mounting medium (Life Technologies). Images were collected using a 63x oil-immersion objective on a Zeiss LSM800 confocal microscope and processed with airyscan. Coverslips were used to quantify colocalization as described below.
Venkatesh H.S., Morishita W., Geraghty A.C., Silverbush D., Gillespie S.M., Arzt M., Tam L.T., Espenel C., Ponnuswami A., Ni L., Woo P.J., Taylor K.R., Agarwal A., Regev A., Brang D., Vogel H., Hervey-Jumper S., Bergles D.E., Suvà M.L., Malenka R.C, & Monje M. (2019). Electrical and synaptic integration of glioma into neural circuits. Nature, 573(7775), 539-545.
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