- PA5-18822 - Invitrogen Antibodies YWHAQ antibody

Parachini-Winter C, Bracha S, Ramsey SA, Yang L, Ho E, Leeper HJ, Curran KM
Journal of veterinary internal medicine 2020 Sep;34(5):2036-2047

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of 14-3-3 theta was performed using HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with 14-3-3 theta Goat Polyclonal Antibody (Product # PA5-18822) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Rabbit anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11078) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the composite image showing cytoplasmic localization of 14-3-3 theta in HeLa cells . Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification..

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of 14-3-3 theta was performed using HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with 14-3-3 theta Goat Polyclonal Antibody (Product # PA5-18822) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Rabbit anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11078) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the composite image showing cytoplasmic localization of 14-3-3 theta in HeLa cells . Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification..

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 2 Immunoblots of lymph node samples for 7 dogs (d) with naive multicentric lymphoma before (D0) and after (D7) supplementation with sulforaphane. The proteins HIP, A, and 14-3-3-theta, B, were detected in all samples (top rows). The immunoblots for beta-actin (second rows) were used to normalize the band volume of HIP and 14-3-3-theta. The normalized expression ratio at D7/D0 is the ratio of the normalized band volume at D7 vs D0 for each dog (third rows). For comparison, the corresponding fold change at D7 vs D0 previously determined by LC-MS/MS is also annotated (bottom rows). FC D7/D0: Fold change at D7 vs D0 (mass spectrometry); NA: Not applicable; NER D7/D0: Normalized expression ratio at D7 vs D0 (immunoblot)

PA5-18822