Histone H1.2 Recombinant Rabbit Monoclonal Antibody [JU43-48]
cat.: ET1706-26
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, ChIP
Clonality: Monoclonal
Clone number: JU43-48
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: 21 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Histone H12 aa 1-50 / 213.
Positive control: Hela cell lysate, 293 cell lysate, MCF-7 cell lysate, rat liver tissue lysate, mouse lung tissue lysate, Hela, PC-3M, SK-Br-3, mouse colon tissue, human kidney tissue, rat brain tissue, human lung carcinoma tissue.
Subcellular location: Nucleus. Chromosome.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  ChIP
1:500-1:2,000
1:50-1:100
1:50-1:100
1:50-1:200
Use 0.5~2 μg for 25 μg of chromatin.
Uniprot #: SwissProt: P16403 Human | P15864 Mouse | P15865 Rat
Alternative names: H1 histone family member 2 H1.a H12_HUMAN H1F2 H1s-1 HIST1H1C Histone 1 H1c Histone cluster 1 H1c Histone H1.2 Histone H1c Histone H1d Histone H1s-1 MGC3992
Images
ET1706-26_1.jpg Fig1: Western blot analysis of Histone H1.2 on different lysates with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/500 dilution.

Lane 1: Hela cell lysate
Lane 2: 293 cell lysate
Lane 3: MCF-7 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 21 kDa
Observed band size: 25 kDa

Exposure time: 2 minutes;

15% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1706-26) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 5,000 dilution was used for 1 hour at room temperature.
ET1706-26_2.jpg Fig2: Western blot analysis of Histone H1.2 on different lysates with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/500 dilution.

Lane 1: rat liver tissue lysate
Lane 2: mouse lung tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 21 kDa
Observed band size: 30 kDa

Exposure time: 2 minutes;

15% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1706-26) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 5,000 dilution was used for 1 hour at room temperature.
ET1706-26_3.jpg Fig3: Immunocytochemistry analysis of Hela cells labeling Histone H1.2 with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-26_4.jpg Fig4: Immunocytochemistry analysis of PC-3M cells labeling Histone H1.2 with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-26_5.jpg Fig5: Immunocytochemistry analysis of SK-Br-3 cells labeling Histone H1.2 with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI.
ET1706-26_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-26) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1706-26_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-26) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1706-26_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-26) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1706-26_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-26) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1706-26_10.jpg Fig10: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Histone H1.2 (ET1706-26) or 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.
ET1706-26_11.jpg Fig11: Immunocytochemistry analysis of C6 cells labeling Histone H1.2 with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1706-26_12.jpg Fig12: Immunocytochemistry analysis of NIH/3T3 cells labeling Histone H1.2 with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H1.2 antibody (ET1706-26) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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