Histone H3 (tri methyl K4) Recombinant Rabbit Monoclonal Antibody [PSH25-68]
cat.: HA724555
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Green monkey
Applications: WB, IF-Cell, IHC-P, FC(Intra), ChIP
Clonality: Monoclonal
Clone number: PSH25-68
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 15 kDa
Isotype: IgG
Positive control: HeLa (Human cervical adenocarcinoma cell) cell lysate, U-2 OS (Human osteosarcoma cell) cell lysate, NIH/3T3 (Mouse fibroblast) cell lysate, C2C12 (Mouse myoblast) cell lysate, C6 (Rat glioma cell) cell lysate, PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate, COS-1 (African green monkey kidney fibroblast) cell lysate.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC(Intra)
  ChIP
1:2,000
1:500
1:50-1:100
1:1,000
Use 0.5~2 μg for 25 μg of chromatin.
Uniprot #: SwissProt: P68431 Human | P68433 Mouse | Q6LED0 Rat
Alternative names: H3 histone family member E pseudogene H3 histone family, member A H3/A H31_HUMAN H3F3 H3FA Hist1h3a HIST1H3B HIST1H3C HIST1H3D HIST1H3E HIST1H3F HIST1H3G HIST1H3H HIST1H3I HIST1H3J HIST3H3 histone 1, H3a Histone cluster 1, H3a Histone H3 3 pseudogene Histone H3.1 Histone H3/a Histone H3/b Histone H3/c Histone H3/d Histone H3/f Histone H3/h Histone H3/i Histone H3/j Histone H3/k Histone H3/l H3K4me3
Images
HA724555_1.JPG Fig1: Western blot analysis of Histone H3 (tri methyl K4) on different lysates with Rabbit anti-Histone H3 (tri methyl K4) antibody (HA724555) at 1/2,000 dilution.

Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: U-2 OS (Human osteosarcoma cell) cell lysate
Lane 3: NIH/3T3 (Mouse fibroblast) cell lysate
Lane 4: C2C12 (Mouse myoblast) cell lysate
Lane 5: C6 (Rat glioma cell) cell lysate
Lane 6: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate
Lane 7: COS-1 (African green monkey kidney fibroblast) cell lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 1 minute 30 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724555, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 15 kDa
Observed band size: 15 kDa
HA724555_2.jpg Fig2: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: HeLa (Human cervical adenocarcinoma cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA724555, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA724555_3.jpg Fig3: Application: Immunocytochemistry (IF-cell)

Species: Mouse
Sample: NIH/3T3 (Mouse fibroblast)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA724555, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA724555_4.jpg Fig4: Application: Immunocytochemistry (IF-cell)

Species: Rat
Sample: C6 (Rat glioma cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA724555, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA724555_5.jpg Fig5: Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Testis
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724555, 1/100, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724555_6.jpg Fig6: Application: Immunohistochemistry (IHC-P)

Species: Rat
Tissue: Testis
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724555, 1/100, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724555_7.jpg Fig7: Application: Flow Cytometry (Intra)

Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: HA724555 (1/1,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.

HA724555_8.jpg Fig8: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H3 (tri methyl K4) (HA724555) / Competitor's antibody / Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.