Alexa fluor 568 goat anti guinea pig

Manufactured by Thermo Fisher Scientific

Alexa Fluor 568 goat anti-guinea pig is a fluorescent secondary antibody used in immunofluorescence assays. It is a conjugate of the Alexa Fluor 568 dye and an antibody raised in goat against guinea pig immunoglobulins.

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Close spelling variants of this product

Alexa Fluor 568 (1190 citations) Alexa 568 (337 citations) Alexa Fluors (21 citations) Anti-goat Alexa-Fluor 568 (6 citations) Alexa Fluor 568 goat (5 citations) Alexa Fluor 568-labeled goat anti-guinea pig IgG antibody (2 citations)

4 protocols using alexa fluor 568 goat anti guinea pig

Immunofluorescence Staining of Cellular Markers

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Cells cultured on glass coverslip were fixed with 4% PFA, treated with 0.1 M glycine in PBS for 30 min, followed by permeabilization with 0.2% saponin in PBS for 30 min and blocked with 5% goat or donkey serum and 0.05% BSA in 0.04% saponin containing PBS [64]. Coverslips were incubated overnight with primary antibodies at 4ºC and then with secondary antibodies in the blocking solution for 2 h at room temperature. Nuclei were stained with DAPI.
Primary antibodies include: SQSTM1 (1:200; Progen, GP62-C), TUBB3/Tuj1 (1:500; Biolegend, 801202), CTSL (1:250, R&D Systems, AF-1515), LAMP1 (1:200; Abcam, 24170). Secondary antibodies: Alexa Fluor 488 goat anti-rabbit (A11034), Alexa Fluor 546 anti-rabbit (A11035), Alexa Fluor 546 goat anti-mouse (A11030), Alexa Fluor 568 goat anti-guinea pig (A11075), Alexa Fluor 633 goat anti-mouse (A21052) and Alexa Fluor 546 donkey anti-goat (A11056) were from Invitrogen.

Sarkar C., Jones J.W., Hegdekar N., Thayer J.A., Kumar A., Faden A.I., Kane M.A, & Lipinski M.M. (2019). PLA2G4A/cPLA2-mediated lysosomal membrane damage leads to inhibition of autophagy and neurodegeneration after brain trauma. Autophagy, 16(3), 466-485.

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Immunofluorescence Staining with Alexa Fluor Antibodies

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Alexa Fluor secondary antibodies (Invitrogen) were used at a dilution of 1:1,000.
Alexa Fluor 488 goat anti-mouse (#A11029), Alexa Fluor 488 goat anti-rabbit (#A11034), Alexa Fluor 488 goat anti-guinea pig (#A11073), Alexa Fluor 488 goat anti-chicken IgY (#A11039), Alexa Fluor 555 goat anti-mouse (#A21422), Alexa Fluor 555 goat anti-rabbit (#A21428), Alexa Fluor 568 goat anti-guinea pig (#A11075), Alexa Fluor 647 goat anti-mouse (#A21236), Alexa Fluor 647 goat anti-rabbit (#A21245). Only cross-adsorbed secondary antibodies were used in this study to eliminate the risk of cross-reactivity.
F-Actin was stained with phalloidin conjugated to Rhodamine (Invitrogen, Cat. #R415, 1:500 dilution).

Chen J, & St Johnston D. (2022). De novo apical domain formation inside the Drosophila adult midgut epithelium. eLife, 11, e76366.

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Immunolabeling of Hippocampal Slices

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Hippocampal slices were washed 3 times with 0.01 M phosphate buffer saline (PBS) and fixed in 4% paraformaldehyde (PFA) for 1 h at room temperature (RT). At the end of fixation, the slices were washed 3 times with PBS and then stored at 4 °C in PBS containing 0.05% sodium azide until use. Prior to immunolabelling, slices were washed 3 times with PBS for 10 min, then incubated overnight with 1% Triton X-100 in PBS at 4 °C, followed by a blocking stage using 20% bovine serum albumin (BSA) diluted in PBS (0.01 M) and containing 0.1% Triton (PBS-T) for 3 h at RT. Slices were then incubated overnight with primary antibody diluted in PBS-T and 1% normal goat serum (NGS): rabbit anti-NeuN antibody (1: 200, Cell Signalling Technology, D4G4O # 24,307), guinea pig anti-GLT1 antibody (1:5000, Merck Millipore Chemicon International, ab1783), rabbit anti-GLAST antibody (1: 150, Abcam, ab416), or rabbit anti-GS (1:5000, Abcam, ab49873). Following washes in PBS-T, slices were incubated in appropriate secondary antibodies for 3 h at RT, namely goat anti-guinea pig Alexa-fluor 568 or goat anti-rabbit Alexa-Fluor 647 (both at 1:500, Invitrogen), in addition to the nuclear chromatin dye Hoechst 33,342 (1:500, Fisher, 11,544,876). Lastly, slices were washed in PBS-T (0.1%) and mounted in Fluoromount G (Invitrogen—REF 00–4958-02).

Ferreira R.S., Ribeiro P.R., Silva J.H., Hoppe J.B., de Almeida M.M., de Lima Ferreira B.C., Andrade G.B., de Souza S.B., Ferdandez L.G., de Fátima Dias Costa M., Salbego C.G., Rivera A.D., Longoni A., de Assis A.M., Pieropan F., Moreira J.C., Costa S.L., Butt A.M, & da Silva V.D. (2023). Amburana cearensis seed extract stimulates astrocyte glutamate homeostatic mechanisms in hippocampal brain slices and protects oligodendrocytes against ischemia. BMC Complementary Medicine and Therapies, 23, 154.

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Immunofluorescent Staining of Pancreatic Islets

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Islets were fixed overnight at 4°C in paraformaldehyde before application of primary antibodies against ADCY5/6 (cat. no. ab66037; Abcam) (34 (link)) and either guinea pig anti-insulin 1:200 or mouse anti-glucagon 1:1,000 (both DAKO). Revelation was performed with goat anti-rabbit Alexa-Fluor 488 and either goat anti–guinea pig Alexa-Fluor 568 or goat anti-mouse Alexa-Fluor 568 antibodies (both 1:500, Invitrogen). Images were acquired as previously described (27 (link)).

Hodson D.J., Mitchell R.K., Marselli L., Pullen T.J., Gimeno Brias S., Semplici F., Everett K.L., Cooper D.M., Bugliani M., Marchetti P., Lavallard V., Bosco D., Piemonti L., Johnson P.R., Hughes S.J., Li D., Li W.H., Shapiro A.M, & Rutter G.A. (2014). ADCY5 Couples Glucose to Insulin Secretion in Human Islets. Diabetes, 63(9), 3009-3021.

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