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Cy5 conjugated anti rabbit igg

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Cy5-conjugated anti-rabbit IgG is a laboratory reagent used in immunoassays and other immunochemical techniques. It is a secondary antibody that binds to rabbit immunoglobulin G (IgG) and is conjugated with the fluorescent dye Cy5. This product can be used to detect and visualize rabbit IgG in samples.

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Lab products found in correlation

Cy3 conjugated anti mouse igg, by Jackson ImmunoResearch (3 mentions) Alexa 488 conjugated anti rat igg, by Thermo Fisher Scientific (2 mentions) Rat anti gfp, by Nacalai Tesque (2 mentions) Rabbit anti cux1, by Santa Cruz Biotechnology (2 mentions) Tcs sp5, by Leica (2 mentions) Mca342r, by Bio-Rad (1 mentions) Ab15323, by Abcam (1 mentions) Guinea pig anti insulin, by Abcam (1 mentions) Goat anti pdx1, by Abcam (1 mentions) Cy3 conjugated anti goat igg, by Jackson ImmunoResearch (1 mentions) Rabbit anti glucagon, by Abcam (1 mentions) 1 4 diazabicyclo 2.2.2 octane, by Merck Group (1 mentions) Normal goat serum (ngs), by Vector Laboratories (1 mentions) Gf090r, by Nacalai Tesque (1 mentions) A11006, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Anti α tubulin, by Merck Group (1 mentions) Anti grp94, by Stressgen (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Anti calnexin, by Stressgen (1 mentions)

8 protocols using cy5 conjugated anti rabbit igg

1

Dual Immunofluorescence Staining of M1 and M2 Microglia

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Mouse monoclonal antibodies against CD163 (1:100; MCA342R, Bio-Rad, Hercules, CA, USA) and rabbit polyclonal antibodies against inducible nitric oxide synthase (iNOS; 1:100; ab15323, Abcam, Cambridge, UK) were used as primary antibodies to label M2 microglia/macrophages and M1 microglia/macrophages, respectively. Cy3-conjugated anti-mouse IgG (10 µg/ml; Jackson ImmunoResearch Laboratories, West Grove, PA, USA) and Cy5-conjugated anti-rabbit IgG (10 µg/ml; Jackson ImmunoResearch Laboratories) were used as secondary antibodies, and DyLight 488-conjugated Lycopersicon Esculentum (Tomato) lectin (Vector Laboratories, Burlingame, CA, USA) was used with 4′,6-diamidino-2-phenylindole (DAPI) to label microglia.
Asano K., Nakamura T, & Funakoshi K. (2022). Early mobilization in spinal cord injury promotes changes in microglial dynamics and recovery of motor function. IBRO Neuroscience Reports, 12, 366-376.
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2

Immunofluorescence Visualization of Cux1 and GFP

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After the real-time luciferase assay, the slices were fixed for several hours with 4% paraformaldehyde in 0.1 M PBS. The slices were incubated at 4°C overnight with rabbit anti-Cux1 (1:250; Santa Cruz Biotechnology) and rat anti-GFP (1:1,000; nacalai tesque). After extensive washes, the signals were visualized with Alexa 488-conjugated anti-rat IgG (1:500; Invitrogen) and Cy5-conjugated anti-rabbit IgG (1:250; Jackson ImmunoResearch). The samples were embedded with 80% glycerol containing DAPI and DABCO, and observed by confocal microscopy through a 20× objective lens (Leica, TCS-SP5).
Miyasaka Y, & Yamamoto N. (2021). Neuronal Activity Patterns Regulate Brain-Derived Neurotrophic Factor Expression in Cortical Cells via Neuronal Circuits. Frontiers in Neuroscience, 15, 699583.
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3

Immunohistochemical Analysis of Pancreatic Islets

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Pancreas sections were prepared from MT-tg (n = 5) and WT male mice (n = 5) at 8 weeks of age. At least three slides from each mouse were examined. Immunostaining was conducted using routine procedures for paraffin embedding and sectioning as described previously [32 (link)]. Briefly, sections were deparaffinized, rehydrated, pretreated with target antigen retrieval solution, washed with PBS, blocked with serum, and incubated with primary antibodies [guinea pig anti-insulin (Abcam, Cambridge, MA, USA), diluted to a ratio of 1:50; mouse anti-horse MT (Dako, Carpinteria, CA, USA), diluted to 1:50, or rabbit anti-glucagon (Abcam), diluted to 1:50; goat anti-PDX1, diluted to 1:50 (Abcam)] at 4°C overnight. After incubation, immune complexes were detected by double-labeling immunohistochemistry, incubation with fluorescein isothiocyanate (FITC)-labeled anti-guinea pig IgG, and Cy3-conjugated anti-mouse IgG or Cy5-conjugated anti-rabbit IgG and Cy3-conjugated anti-goat IgG (1:200; all from Jackson ImmunoResearch Laboratories, West Grove, PA, USA).
Chen S., Han J, & Liu Y. (2015). Dual Opposing Roles of Metallothionein Overexpression in C57BL/6J Mouse Pancreatic β-Cells. PLoS ONE, 10(9), e0137583.
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4

Immunostaining of Cortical Neurons

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Cortical neurons were fixed at room temperature for 10 min in 4% paraformaldehyde/PBS. They were then permeabilized and blocked for 15 min in buffer G, composed of 5% normal goat serum (Vector Labs) and 0.1% Triton X-100 in PBS. The cells were then incubated overnight at 4 °C with the primary antibody in buffer G. The antibodies used are as follows: rabbit polyclonal anti-CREB antibody (GeneTex, 1:100), mouse monoclonal anti-CREB antibody (86B10, Cell Signal Technology, 1:100), rabbit monoclonal anti-c-Fos antibody (9F6, Cell Signal Technology, 1:200) and rat monoclonal anti-GFP antibody (GF090R, Nacalai Tesque, 1:1000). For visualization, the cultures were further incubated at RT for 2 h in buffer G containing the following secondary antibodies: Alexa 488-conjugated anti-rat IgG (A11006, Life Technologies, 1:400), Cy3-conjugated anti-mouse IgG (AP192C, Millipore, 1:200), Cy3-conjugated anti-rabbit IgG (AP182C, Millipore, 1:400) or Cy5-conjugated anti-rabbit IgG (711-175-166, Jackson ImmunoResearch, 1:200). Nuclei were stained with 0.1% 4′,6-diamidino-2-phenylindole (DAPI, Sigma) in a mounting medium containing 50% glycerol and 2.3% 1,4-diazabicyclo[2.2.2]octane (Sigma) in 50 mM Tris–HCl (pH 8.0).
Tsunematsu H., Uyeda A., Yamamoto N, & Sugo N. (2017). Immunocytochemistry and fluorescence imaging efficiently identify individual neurons with CRISPR/Cas9-mediated gene disruption in primary cortical cultures. BMC Neuroscience, 18, 55.
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5

Antibody Sources for GFP, RFP, and PrP-A

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Rabbit antisera for GFP and RFP were made as described (Stefanovic and Hegde, 2007 ) and rabbit antisera for Tmp21 was made by immunizing rabbits with a peptide corresponding to the cytoplasmic tail (CLRRFFKAKKLIE) conjugated to keyhole limpet hemacyanin via the terminal cysteine. The PrP-A antibody was previously described (Rane et al., 2004 (link)). The antibody against TCR-α, A2B4, (gift from Carolyn Suzuki) was previously described (Suzuki et al., 1991 (link)). CHOP and XBP-1 antibodies were gifts from Linda Hendershot.
Purchased antibodies included anti-calnexin (StressGen SPA-860), anti-Grp94 (StressGen SPA-850), anti-ERGIC53 (Santa Cruz SC-66880), anti-alpha tubulin (Sigma T6199), anti-GAPDH (Santa Cruz SC-20357), Cy5-conjugated anti-rabbit IgG (Jackson ImmunoResearch 111-175-003) and Alexa 488 goat anti-mouse Ab (Invitrogen A11029).
Satpute-Krishnan P., Ajinkya M., Bhat S., Itakura E., Hegde R.S, & Lippincott-Schwartz J. (2014). ER Stress-Induced Clearance of Misfolded GPI-Anchored Proteins via the Secretory Pathway. Cell, 158(3), 522-533.
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6

Immunohistochemical and Ultrastructural Analysis

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Immunohistochemistry was carried out on 10 μm sections as described in detail elsewhere (14 (link), 15 (link)). Briefly, non-specific binding sites were quenched with 5% bovine serum albumin; sections were then incubated with rabbit anti-entactin antibody (Abcam, Cambridge, UK) followed by Cy5-conjugated anti-rabbit IgG (Jackson Immunoresearch, West Grove, PA) for 45 minutes. Sections were counterstained with TRITC-conjugated phalloidin (Sigma-Aldrich) and analysed with a Zeiss LSM 510 confocal microscope. For TEM analysis, samples were processed according to standard procedure (16 (link)) and examined with a Philips 201 electron microscope.
Man A.L., Gicheva N., Regoli M., Rowley G., De Cunto G., Wellner N., Bassity E., Gulisano M., Bertelli E, & Nicoletti C. (2016). CX3CR1+ cell-mediated Salmonella-exclusion protects the intestinal mucosa during the initial stage of infection. Journal of immunology (Baltimore, Md. : 1950), 198(1), 335-343.
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7

Immunocytochemistry Protocol for Integrin Internalization

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For immunocytochemistry, cells were seeded on Poly-L-lysine (Sigma, P7407) coated 24 well Chamber slides, grown until 80~90% confluency and fixed with 4% PFA for 15 min. Fixed monolayer cells were washed twice with ice-cold PBS and incubated with 0.25% Triton X-100 in PBS for 15 min for permeabilization. Cells were incubated with 1% BSA for 30 min for blocking of non-specific binding, followed by primary antibodies at 4°C overnight. After 2 washes in ice-cold PBS, primary antibodies were detected with cy3-conjugated anti-mouse IgG and cy5-conjugated anti-rabbit IgG (Jackson Laboratories), followed by DAPI staining. Confocal images were taken with a Zeiss LSM 800 and LSM780 confocal microscope using Zeiss software (Carl Zeiss Micro Imaging, Jena, Germany). The assessment of integrin internalization was performed as described in supplementary material and method.
Jeong H., Lim K.M., Kim K.H., Cho Y., Lee B., Knowles B.C., Roland J.T., Zwerner J.P., Goldenring J.R, & Nam K.T. (2019). Loss of Rab25 promotes the development of skin squamous cell carcinoma through the dysregulation of integrin trafficking. The Journal of pathology, 249(2), 227-240.
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8

Immunohistochemical analysis of Cux1 and GFP

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After the real-time luciferase assay, the slices were fixed for several hours with 4% paraformaldehyde in 0.1 M PBS. The slices were incubated at 4°C overnight with rabbit anti-Cux1 (1:250; Santa Cruz Biotechnology) and rat anti-GFP (1:1000; nacalai tesque). After extensive washes, the signals were visualized with Alexa 488-conjugated anti-rat IgG (1:500; invitrogen) and Cy5-conjugated anti-rabbit IgG (1:250; Jackson ImmunoResearch). The samples were embedded with 80% glycerol containing DAPI and DABCO, and observed by confocal microscopy through a 20× objective lens (Leica, TCS-SP5).
Miyasaka Y., & Yamamoto N. (2021). Neuronal activity patterns regulate BDNF expression in cortical neurons via synaptic connections and calcium signaling.
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