bs-0349R [Primary Antibody]
Histone H3 Polyclonal Antibody
www.biossusa.com
[email protected]
800.501.7654 [DOMESTIC]
+1.781.569.5821 [INTERNATIONAL]
DATASHEET

Host: Rabbit

Target Protein: Histone H3

Immunogen Range: 71-136/136


Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 3020

Swiss Prot: P84243

Source: KLH conjugated synthetic peptide derived from human Histone H3

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

Storage: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Background:

Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.

Size: 100ul

Concentration: 1ug/ul

Applications: WB(1:300-5000)
ELISA(1:500-1000)
FCM(1:20-100)
IHC-P(1:200-400)
IHC-F(1:100-500)
IF(IHC-P)(1:50-200)
IF(IHC-F)(1:50-200)
IF(ICC)(1:50-200)
ICC(1:100-500)

Predicted Molecular Weight: 15


Cross Reactive Species: Human
Mouse
Rat
Others

Predicted Cross Reactive Species: Cow
Pig
Rabbit
Drosophila

For research use only. Not intended for diagnostic or therapeutic use.

PRODUCT SPECIFIC PUBLICATIONS
  • Liu, Donghui, et al. ʺNonenzymatic glycation of high‐density lipoprotein impairs its anti‐inflammatory effects in innate immunity.ʺ Diabetes/Metabolism Research and Reviews 28.2 (2012): 186-195.Read more>>
  • Wang, Chunqiang, Wei Ma, and Yuhong Su. ʺNF-κB Pathway Contributes to Cadmium-Induced Apoptosis of Porcine Granulosa Cells.ʺ Biological trace element research (2013): 1-8.Read more>>
  • Duan, Chao, et al. "Oestrogen receptor‐mediated expression of Olfactomedin 4 regulates the progression of endometrial adenocarcinoma." Journal of Cellular and Molecular Medicine (2014).Read more>>
  • Tang, Z-X., et al. "Selective antegrade cerebral perfusion attenuating the TLR4/NF-κB pathway during deep hypothermia circulatory arrest in a pig model." Cardiology 128.3 (2014): 243-250.Read more>>
  • Qian, Yi, et al. "Silver Nanoparticle-Induced Hemoglobin Decrease Involves Alteration of Histone 3 Methylation Status." Biomaterials (2015).Read more>>
  • Zhao, Hongyu, et al. "Glycyrrhizic Acid Attenuates Sepsis-Induced Acute Kidney Injury by Inhibiting NF-κB Signaling Pathway." Evidence-Based Complementary and Alternative Medicine (2015).Read more>>
  • Shen, Haitao, et al. "Epigallocatechin-3-gallate alleviates paraquat-induced acute lung injury and inhibits upregulation of toll-like receptors." Life Sciences (2016).Read more>>
  • Dong, Wei-tao, et al. "iTRAQ proteomic analysis of the interactions between Bombyx mori nuclear polyhedrosis virus and silkworm." Journal of Proteomics (2017).Read more>>
  • Joy, Marion, et al. "The Myocardin-related transcription factor MKL co-regulates the cellular levels of two profilin isoforms." Journal of Biological Chemistry (2017): jbc-M117.Read more>>
  • Zhang et al. Uncoupling Protein 2 Inhibits Myointimal Hyperplasia in Preclinical Animal Models of Vascular Injury. (2017) J.Am.Heart.Assoc. 6 Read more>>
  • Zhou et al. Epigallocatechin-3-gallate inhibits proliferation and migration of human colon cancer SW620 cells in vitro. (2012) Acta.Pharmacol.Sin. 33:120-6Read more>>
  • Yang et al. Celastrol Attenuates Multiple Sclerosis and Optic Neuritis in an Experimental Autoimmune Encephalomyelitis Model. (2017) Front.Pharmacol. 8:44Read more>>
  • Peng et al. Overexpressing modified human TRβ1 suppresses the proliferation of breast cancer MDA-MB-468 cells. (2018) Oncol.Lett. 16:785-792Read more>>
  • Lin,et al.The CXCL12/CXCR4 axis promotes migration, invasion and EMT in human papillary thyroid carcinoma B-CPAP cells via NF-κB signaling.(2018) Biochemistry and Cell Biology. :.Read more>>
  • Sogorkova,et al.Optimization of cell growth on palmitoyl-hyaluronan knitted scaffolds developed for tissue engineering applications.(2018) Journal of Biomedical Materials Research. Part A. 106:1488-1499.Read more>>
  • Cui Y et al. Loganin prevents BV‐2 microglia cells from Aβ1‐42‐induced inflammation via regulating TLR4/TRAF6/NF‐κB axis.(2019) Cell Biol Int.Read more>>
  • Li L et al. Microtubule associated protein 4 phosphorylation leads to pathological cardiac remodeling in mice.(2018)EBioMedicine Oct 13Read more>>
  • Si J et al. Linagliptin protects rat carotid artery from balloon injury and activates the NRF2 antioxidant pathway.(2018) Exp Anim. Oct 23.Read more>>
  • Wang J et al. PIM1 inhibitor SMI-4a attenuated lipopolysaccharide-induced acute lung injury through suppressing macrophage inflammatory responses via modulating p65 phosphorylation. Int Immunopharmacol. 2019 Aug;73:568-574.Read more>>
  • Ji X et al. Histone modification in the lung injury and recovery of mice in response to PM2.5 exposure.. Chemosphere. 2019 Apr;220:127-136. Read more>>
  • Wang L et al. PHD2 exerts anti-cancer and anti-inflammatory effects in colon cancer xenografts mice via attenuating NF-κB activity. Life Sci. 2020 Feb 1;242:117167. Read more>>
  • Huina Liu. et al. LncRNA, PLXDC2\OT promoted the osteogenesis potentials of MSCs by inhibiting the deacetylation function of RBM6/SIRT7 complex and OSX specific isoform. 2021 Mar 08Read more>>
  • Wang Li. et al. Trilobatin Induces Apoptosis and Attenuates Stemness Phenotype of Acquired Gefitinib Resistant Lung Cancer Cells via Suppression of NF-B Pathway. 2021 Apr 16Read more>>
  • Zhuohong Li. et al. Anlotinib exerts anti-cancer efficiency on lung cancer stem cells in vitro and in vivo through reducing NF-B activity. 2021 May 06Read more>>
  • Zhao-Ming Xiao. et al. SMARCC1 Suppresses Tumor Progression by Inhibiting the PI3K/AKT Signaling Pathway in Prostate Cancer. Front Cell Dev Biol. 2021; 9: 678967Read more>>
  • Zhiwei Wang. et al. Dihydroartemisinin triggers ferroptosis in primary liver cancer cells by promoting and unfolded protein response?induced upregulation of CHAC1 expression. Oncol Rep. 2021 Nov;46(5):1-14Read more>>
  • Zheng Hao. et al. Decreased Expression of Programmed Death Ligand-L1 by Seven in Absentia Homolog 2 in Cholangiocarcinoma Enhances T-CellMediated Antitumor Activity. Front Immunol. 2022 Jan;0:138Read more>>
  • Huiling Mao. et al. Leucine protects bovine intestinal epithelial cells from hydrogen peroxide-induced apoptosis by alleviating oxidative damage. J SCI FOOD AGR. 2022 May 05Read more>>
  • Fanchun Zeng. et al. Antagonizing exosomal miR-18a-5p derived from prostate cancer cells ameliorates metastasis-induced osteoblastic lesions by targeting Hist1h2bc and activating Wnt/_-catenin pathway. GENES DIS. 2022 JulRead more>>
  • Minkai Song. et al. circIFNGR2 Regulating Ankylosing Spondylitis Associated Inflammation through Macrophage Polarization. ISCIENCE. 2023 Jul;:107325Read more>>
  • Dongliu Luo. et al. Cannabidiol alleviates perfluorooctane sulfonate-induced macrophage extracellular trap mediate inflammation and fibrosis in mice liver. ECOTOX ENVIRON SAFE. 2023 Sep;263:115374Read more>>
VALIDATION IMAGES

Paraformaldehyde-fixed, paraffin embedded human colon cancer; 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°C for 30min; Antibody incubation with HIST3H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.


Formalin-fixed and paraffin embedded rat brain labeled with Rabbit Anti Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 followed by conjugation to the secondary antibody and DAB staining


Formalin-fixed and paraffin embedded mouse intestine labeled with Rabbit Anti Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 followed by conjugation to the secondary antibody and DAB staining


Formalin-fixed and paraffin-embedded rat brain labeled with Anti-Histone H3 Polyclonal Antibody, Unconjugated(bs-0349R) 1:200, overnight at 4°C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C.


Mouse splenocytes probed with Rabbit Anti-Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) (green) at 1:50 for 30 minutes followed by a FITC conjugated secondary antibody compared to unstained cells (blue), secondary only(light blue), and isotype control(orange).


K562 Cell lysates probed with Histone H3 Polyclonal Antibody, unconjugated (bs-0349R) at 1:300 overnight at 4°C followed by a conjugated secondary antibody at 1:10000 for 60 minutes at 37°C.


Paraformaldehyde-fixed, paraffin embedded Rat liver; 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Rat colon; 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Human laryngeal cancer; 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Human colon; 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Mouse intestine; 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Rat pancreas; 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Mouse pancreas; 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Mouse testis; 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Rat testis; 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Mouse 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Human 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Rat testis; 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°C for 30min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded (mouse liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Paraformaldehyde-fixed, paraffin embedded (mouse testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Paraformaldehyde-fixed, paraffin embedded (mouse kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Paraformaldehyde-fixed, paraffin embedded (rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Paraformaldehyde-fixed, paraffin embedded (human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Paraformaldehyde-fixed, paraffin embedded (rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Paraformaldehyde-fixed, paraffin embedded (human liver ); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Paraformaldehyde-fixed, paraffin embedded (human brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.


Lane 1: Mouse Raw264.7 cell lysates; Lane 2: Human Hela cell lysates; Lane 3: Human SH-SY5Y cell lysates; Lane 4: Human Molt-4 cell lysates ; Lane 5: Human THP-1 cell lysates ; Lane 6: Human 293T cell lysates probed with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:50000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.


Paraformaldehyde-fixed, paraffin embedded (rat liver); 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°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control) ) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.


Paraformaldehyde-fixed, paraffin embedded (mouse liver); 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°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control) ) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.


Paraformaldehyde-fixed, paraffin embedded (mouse testis); 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°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control) ) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.


Paraformaldehyde-fixed, paraffin embedded (rat testis); 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°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control) ) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.


25 ug total protein per lane of various lysates (see on figure) probed with Histone H3 polyclonal antibody, unconjugated (bs-0349R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.