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Recombinant Antibodies

Recombinant Antibodies


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Western blot was performed on whole cell (25 \u00b5g, lane 1) and histone extracts (15 \u00b5g, lane 2) from HeLa cells, and on 1 \u00b5g of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Bioss antibody against H3K9me3 (Cat. No. bs-53114R). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk.
HeLa cells were stained with the Bioss antibody against H3K9me3 (bs-53114R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K9me3 antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The left panel shows staining of the nuclei with DAPI. A merge of both stainings is shown on the right.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against human H3K9me3 (bs-53114R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:30,000.
A Dot Blot analysis was performed to test the cross-reactivity of the antibody against H3K9me3 (Cat. No. bs-53114R) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification 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 modification of interest.

Catalog #1: bs-53114r

Applications: WB, IHC-P, IHC-F, IF, ICC/IF

Reactivity: Human, Mouse, Rat

Conjugate: Unconjugated

25 ug total protein per lane of various lysates (see on figure) probed with ChAT monoclonal antibody, unconjugated (bsm-54807R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ChAT Monoclonal Antibody, Unconjugated(bsm-54807R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ChAT Monoclonal Antibody, Unconjugated(bsm-54807R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.

Catalog #1: bsm-54807r-100ul

Applications: WB, IHC-P, IHC-F, IF(IHC-P)

Reactivity: Mouse, Rat

Conjugate: Unconjugated

Catalog #1: bsm-90294R

Applications: WB

Reactivity: Human, Mouse, Rat

Conjugate: Unconjugated

Catalog #1: bsm-90295R

Applications: WB

Reactivity: Human, Rat

Conjugate: Unconjugated