PA5-51466

antibody from Invitrogen Antibodies

Targeting: H1-0 H1.0, H1F0, H1FV

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-51466

Provider

Invitrogen Antibodies

Product name

Histone 1F0 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: MTENSTDHPK YSDMIVAAIQ AEKNRAGSSR QSIQKYIKSH YKVGENADSQ IKLSIKRLVT TGVLKQTKGV GASGSFRLAK SDEPKKSVAF KKTKKEIKKV ATPKKASK

Concentration

0.05 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Histone 1F0 in Lane 1: Marker (kDa) 250, 130, 100, 70, 55, 35, 25, 15, 10; Lane 2: Human cell line U-2 OS. Samples were probed using a Histone 1F0 Polyclonal Antibody (Product # PA5-51466).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on acid extracts (20 µg lysate) of PC-12 (Lane 1), PC-12 treated with Retinoic acid (100 uM for 12 days) (Lane 2), HEK-293 (Lane 3), MDA-MB-231 (Lane 4), DU-145 (Lane 5), 3T3-L1 (Lane 6), U-2 OS (Lane 7) and A-549 (Lane 8). The blot was probed with Anti-Histone 1F0 Polyclonal Antibody (Product # PA5-51466, 1:500 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 30 kDa band corresponding to Histone 1F0 was observed across the cell lines tested and enhanced upon differentiation with retinoic acid.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent staining of Histone 1F0 in human cell line A-431 using a Histone 1F0 Polyclonal Antibody (Product # PA5-51466) shows localization to nucleus and actin filaments.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Histone 1F0 was performed using 70% confluent log phase U2OS cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Histone 1F0 Rabbit Polyclonal Antibody (Product # PA5-51466) at 5 microgram/mL in 0.1% BSA and incubated overnight at 4 degree 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 SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of Histone 1F0 in human prostate and lymph node tissues using Histone 1F0 Polyclonal Antibody (Product # PA5-51466). Corresponding H1F0 RNA-seq data are presented for the same tissues.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of Histone 1F0 in human prostate using Histone 1F0 Polyclonal Antibody (Product # PA5-51466) shows high expression.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of Histone 1F0 in human lymph node using Histone 1F0 Polyclonal Antibody (Product # PA5-51466) shows low expression as expected.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Enrichment of endogenous Histone H2A.X protein at specific gene loci using Anti-Histone H2A.X Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-Histone H2A.X Polyclonal Antibody (Product # PA1-41004, 3 µg) on sheared chromatin from 2 million U-2 OS cells using the "MAGnify ChIP system" kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over the GAPDH gene (active) and MYOD, SAT2 satellite repeats, SAT Alpha (inactive). A schematic diagram of the GAPDH gene is shown on top of the figure. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.