RECEIVE 25% OFF PUBLISHED ANTIBODIES WITH CODE "PUB25"! Offer valid for US customers and available internationally through distributors.

Contact us at 1.800.501.7654 or info@biossusa.com

MenuSearch

Unconjugated Primary Antibodies



×

Collection Menu
To determine the titer of the antibody, an ELISA was performed using a serial dilution of Bioss antibody directed against human DNMT3A (Cat. No. bs-53128R), crude serum and Flow Through, in antigen-coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:500.
Human embryonic kidney cells (HEK293T) were transiently transfected with an expression vector for GAL4- tagged DNMT3A. Whole cell extracts were immunoprecipitated with 2 \u03bcg of DNMT3A Polyclonal Antibody (Cat. No. bs-53128R). The presence of GAL4-DNMT3A in the non-treated cell extracts and in the immunoprecipitates was demonstrated by western blot analysis with the anti-GAL4 antibody (lane 1 and 2 respectively). Alternatively, GAL4-DNMT3A was immunoprecipitated and western blot analysis was performed with the DNMT3A antibody (diluted 1:2,000) (lane 3).

Catalog #1: bs-53128R

Applications: WB, ELISA, IP, ChIP-seq

Reactivity: Human

Conjugate: Unconjugated

ChIP assays were performed using HeLa cells treated with colcemid, the H3K27me3S28p antibody (bs-53053R) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 10,000 cells. A titration of the antibody consisting of 1, 5 and 10 \u03bcl per ChIP experiment was analysed. Additionally, ChIP was performed after incubation of the antibody with 5 nmol blocking peptide (cat. No. sp-091-050) for 1 hour at room temperature. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active genes c-fos and RPL30 and for the inactive gene MYOD. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
To determine the titer, an ELISA was performed using a serial dilution of the H3K27me3S28p antibody (bs-53053R). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:8,300.
A Dot Blot analysis was performed to test the cross reactivity of the H3K27me3S28p antibody (bs-53053R) with peptides containing other modifications of histone H3 and H4 and unmodified sequences from histone H3. One hundred to 0.2 pmol of the peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the peptide containing the modifications of interest.
Hela cells were stained with the H3K27me3S28p antibody (bs-53053R) and with DAPI. Cells were fixed with formaldehyde, permeabilized with sodium citrate and Triton X100 and blocked with PBS containing 2.5% BSA. Left side: cells were immunofluorescently labelled with the H3K27me3S28p antibody (diluted 1:200 and incubated for 1 hour at room temperature) followed by goat anti-rabbit antibody conjugated to DyLight 488. Right side: the nuclei were stained with DAPI, which specifically labels DNA. Phosphorylation of H3 on serine 28 is increased during late G2 phase and reaches a maximum in metaphase cells. This may explain the different staining intensities of different cells.

Catalog #1: bs-53053R

Applications: WB, ELISA, IP, IF, dot-blot, ChIP-seq

Reactivity: Human

Conjugate: Unconjugated

Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with PHAP1 Polyclonal Antibody, Unconjugated (bs-6083R) at 1:400 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with PHAP Polyclonal Antibody, Unconjugated (bs-6083R) at 1:400 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with PHAP Polyclonal Antibody, Unconjugated (bs-6083R) at 1:400 overnight at 4\u00b0C, DAB staining.
Lane 1: 293T cell lysates probed with PHAP Polyclonal Antibody, Unconjugated (bs-6083R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Catalog #1: bs-6083R

Applications: WB, ELISA, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC), IP

Reactivity: Human, Mouse, Rat

Conjugate: Unconjugated

Catalog #1: bs-4032R

Applications: WB, ELISA, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC), IP

Reactivity: Human

Conjugate: Unconjugated