Cy3 affinipure donkey anti rat igg h l

Manufactured by Jackson ImmunoResearch

Cy3 AffiniPure Donkey Anti-Rat IgG (H+L) is a secondary antibody that binds to rat IgG antibodies. It is labeled with the fluorescent dye Cy3, which can be used for detection and visualization in various immunoassay techniques.

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Lab products found in correlation

Anti myc clone 9b11, by Cell Signaling Technology (1 mentions) Anti eea1, by BD (1 mentions) Anti ha, by Roche (1 mentions) Anti sara, by Proteintech (1 mentions) Anti ha, by Santa Cruz Biotechnology (1 mentions) Tcs sp2 confocal microscope, by Leica (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Glycine, by Merck Group (1 mentions) Alexa fluor 488 affinipure donkey anti mouse igg h l, by Jackson ImmunoResearch (1 mentions) Mouse anti α tubulin t5168, by Merck Group (1 mentions) Vectashield mounting medium, by Vector Laboratories (1 mentions)

Close spelling variants of this product

Anti-rat Cy3 (13 citations) Anti-rat IgG Cy3 (6 citations) Cy3-AffiniPure donkey anti-rat IgG (3 citations) Donkey Anti-Rat IgG (H+L) (3 citations)

2 protocols using cy3 affinipure donkey anti rat igg h l

Antibody Validation and Usage Protocol

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The following commercial antibodies were used: Mouse: anti-EEA1 (BD Biosciences Cat. No. 610457); anti-HA (for immunoblotting) (Santa Cruz Cat. No Sc-7392); anti-myc 9B11 clone (Cell Signalling Technology Cat. No. 2276); anti-His (Sigma Clone His-1; Cat. No. H1029). Rat: anti-HA (for IF) (Roche; clone 3F10). Rabbit: anti-HD-PTP (Proteintech Cat. No.102472-1-AP), anti-endofin (Proteintech Cat. No. 13118-2-AP), anti-SARA (Proteintech Cat. No. 22033-1-AP). The anti-endofin and anti-SARA antibodies were verified by siRNA (Figure S1A) using OnTargetPlus siRNA Smartpool reagents (Dharmacon) for endofin (LQ-020254-01-0005) and SARA (LQ-011939-00-0005), vs a Dharmacon control siRNA. Fluorescent secondary antibodies for IF or for immunoblotting were from Jackson ImmunoResearch Laboratories (PA, USA). Rat-Mouse double labeling IF experiments used Cy3 AffiniPure Donkey Anti-Rat IgG (H+L), immunoadsorbed to prevent cross-reactivity towards mouse IgG (Catalogue 712-165-153), and Alexa488 AffiniPure Donkey Anti-Mouse IgG (H+L), immunoadsorbed to prevent cross-reactivity towards rat IgG (catalogue 715-545-150) (Jackson ImmunoResearch laboratories).

Gahloth D., Levy C., Walker L., Wunderley L., Mould A.P., Taylor S., Woodman P, & Tabernero L. (2017). Structural Basis for Specific Interaction of TGFβ Signaling Regulators SARA/Endofin with HD-PTP. Structure(London, England:1993), 25(7), 1011-1024.e4.

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Immunofluorescence Staining of Cytoskeleton and Mitosis Markers

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Cells at subconfluence were fixed with 4% paraformaldehyde (PFA)/PBS for 10 min at 4°C, washed twice with PBS 1X, and incubated with 3% BSA (Sigma-Aldrich) and 0.5% TritonX-100 for 10 min at RT. The reduction of nonspecific background signal was obtained by incubating cells with 1 M glycine (Sigma-Aldrich) and subsequently with block solution consisting of 0.5% Triton X-100, 3% BSA, and 5% normal donkey serum (Jackson Laboratories Immuno Research) for 30min at RT. The cells were incubated with the following primary antibodies: mouse anti-α tubulin (T5168; 1:500; Sigma-Aldrich) and rat anti-PHH3 (phosphorylated histone H3) (1:100; Abcam, ab10543) for 1 h at RT. After the washes, the cells were incubated with species-specific secondary antibodies Alexa Fluor® 488 AffiniPure Donkey Anti-Mouse IgG (H+L) and Cy3-AffiniPure Donkey Anti-Rat IgG (H+L) (#715-545-150 and #712-165-153, respectively; Jackson Immuno Research), diluted 1:200 for 1 h at RT. Nuclei were then counterstained with TO-PRO-3 Iodide (642/661) (T3605; Life Technologies), and the slides were closed with Vectashield mounting medium (Vector Laboratories). Pictures were acquired using a Leica TCS SP2 confocal microscope.

Taggi M., Kovacevic A., Capponi C., Falcinelli M., Cacciamani V., Vicini E., Canipari R, & Tata A.M. (2022). The activation of M2 muscarinic receptor inhibits cell growth and survival in human epithelial ovarian carcinoma. Journal of cellular biochemistry, 123(9).

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