PA5-38116

antibody from Invitrogen Antibodies

Targeting: HDAC5 FLJ90614, KIAA0600, NY-CO-9

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-38116

Provider

Invitrogen Antibodies

Product name

Anti-Phospho-HDAC5 (Ser259) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Phospho-HDAC5 pSer259 in extracts from HepG2 cells using a Phospho-HDAC5 pSer259 polyclonal antibody (Product # PA5-38116).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on modified whole cell extracts (1% SDS) (30 µg lysate) of IMR32 (Lane 1), IMR32 treated with Ionomycin (0.5 uM for 1 min) (Lane 2), IMR32 treated with Ionomycin (0.5 uM for 5 min) (Lane 3), IMR32 treated with Ionomycin (0.5 uM for 60 min) (Lane 4), Jurkat (Lane 5), Jurkat treated with Ionomycin (0.5 uM for 1 min) (Lane 6), Jurkat treated with Ionomycin (0.5 uM for 5 min) (Lane 7), Jurkat treated with Ionomycin (0.5 uM for 60 min) (Lane 8). The blot was probed with Anti-Phospho-HDAC5 (Ser259) Polyclonal Antibody (Product # PA5-38116, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 122 kDa band corresponding to Phospho-HDAC5 (Ser259) was observed across the cell lines tested and showed treatment dependent difference in RAW 264.7 cell line upon Ionomycin treatment with varying time points.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Phospho-HDAC5 (Ser259) was performed using 70% confluent log phase Jurkat cells treated with Ionomycin. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-HDAC5 (Ser259) Polyclonal Antibody(Product # PA5-38116) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 merged image showing increased expression upon Ionomycin treatment for 5min. Panel e represents the control cells showing weak nuclear staining. Panel f shows decreased staining upon Ionomycin treatment for 60 min. Panel g represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.