ET1706-15

macroH2A.1 Recombinant Rabbit Monoclonal Antibody [JU22-83]

cat.: ET1706-15

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P
Clonality:	Monoclonal
Clone number:	JU22-83

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:	Predicted band size: 39 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human macroH2A1 aa 232-281 / 369.
Positive control:	Hela cell lysate, Mouse placenta tissue lysate, MCF-7 cell lysate, Hela, HepG2, PC-3M, mouse liver tissue, rat liver tissue, human thyroid gland tissue, human placenta tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P	1:500-1:2,000 1:50-1:200 1:50-1:200 1:5,000-1:10,000
Uniprot #:	SwissProt: O75367 Human \| Q9QZQ8 Mouse \| Q02874 Rat
Alternative names:	Core histone macro h2a.1 Core histone macro-H2A.1 H2A histone family member Y H2A.y H2A/y H2AF12M H2AFJ H2afy H2AY_HUMAN Histone H2A.Y Histone macroH2A1 Histone macroH2A1.1 Histone macroH2A1.2 Macroh2a1 MACROH2A1.1 MacroH2A1.2 Medulloblastoma antigen MU MB 50.205 Medulloblastoma antigen MU-MB-50.205 mH2a mH2A1

Images

	Fig1: Western blot analysis of macroH2A.1 on different lysates with Rabbit anti-macroH2A.1 antibody (ET1706-15) at 1/500 dilution. Lane 1: Hela cell lysate (10 µg/Lane) Lane 2: Mouse placenta tissue lysate (20 µg/Lane) Lane 3: MCF-7 cell lysate (10 µg/Lane) Predicted band size: 39 kDa Observed band size: 39 kDa Exposure time: 2 minutes; 10% 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 (ET1706-15) 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 1:200,000 dilution was used for 1 hour at room temperature.
	Fig2: Immunocytochemistry analysis of Hela cells labeling macroH2A.1 with Rabbit anti-macroH2A.1 antibody (ET1706-15) 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-macroH2A.1 antibody (ET1706-15) 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.
	Fig3: Immunocytochemistry analysis of HepG2 cells labeling macroH2A.1 with Rabbit anti-macroH2A.1 antibody (ET1706-15) 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-macroH2A.1 antibody (ET1706-15) 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.

	Fig4: Immunocytochemistry analysis of PC-3M cells labeling macroH2A.1 with Rabbit anti-macroH2A.1 antibody (ET1706-15) 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-macroH2A.1 antibody (ET1706-15) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-macroH2A.1 antibody (ET1706-15) at 1/10,000 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 (ET1706-15) at 1/10,000 dilution for 1 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-macroH2A.1 antibody (ET1706-15) at 1/5,000 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 (ET1706-15) at 1/5,000 dilution for 1 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.

Fig7: Immunohistochemical analysis of paraffin-embedded human thyroid gland tissue with Rabbit anti-macroH2A.1 antibody (ET1706-15) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1706-15) at 1/50 dilution for 1 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.

Fig8: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-macroH2A.1 antibody (ET1706-15) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1706-15) at 1/50 dilution for 1 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.