Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Immunohistochemical analysis of paraffin embedded human prostate hyperplasia tissue using ab150402 showing positive staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Histone H2A.Z (red) with purified ab150402 at a 1/2500 dilution. Cells were fixed with 80% methanol and permeabilized with 0.1% Tween-20. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast tissue sections labeling Histone H2A.Z with purified ab150402 at 1/2000 dilution (1.09 μg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling Histone H2A.Z with ab150402 (unpurified) at 1/250 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Immunofluorescent analysis of HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Histone H2A.Z with ab150402 (unpurified) at 1/100 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Histone H2A.Z with ab150402 (unpurified) at 1/250 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Histone H2A.Z with purified ab150402 at 1/200 dilution (10 μg/ml). Cells were fixed in 100% Methanol. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Immunohistochemical analysis of paraffin embedded human cervical carcinoma tissue using ab150402 (unpurified) showing positive staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Immunohistochemical analysis of paraffin embedded normal human kidney tissue using ab150402 (unpurified) showing positive staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Immunohistochemical analysis of paraffin embedded human ovarian carcinoma tissue using ab150402 (unpurified) showing positive staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ChIP

Lab

ChIP - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab150402 (unpurified) (blue), and 20μl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.

• ChIP

Unknown

ChIP - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol*. Cells were fixed with formaldehyde for 10 minutes.
The ChIP was performed with 25 μg of chromatin, 2 μg of ab150402 (red), and 20 μl of Protein A/G sepharose beads. 2 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers and probes are located in the first kb of the transcribed region.
*http : //www.abcam.com/resources?keywords=X%20ChIP%20protocol

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue sections labeling Histone H2A.Z with purified ab150402 at 1/2000 dilution (1.09 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat cerebrum tissue sections labeling Histone H2A.Z with purified ab150402 at 1 : 2000 dilution (1.09 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Lab

Western blot - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Blocking buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (ab150402) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in 1% SDS Hot lysis method at 15 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates prepared in RIPA lysis method at 15 µg

Lane 4:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates prepared in 1% SDS Hot lysis method at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 14 kDa

Observed band size: 13 kDa

false

• WB

AbReview47197****

Western blot - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

All lanes:

Western blot - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (ab150402) at 1/1000 dilution

Lane 1:

Recombinant Human octamers containing H2A at 1 µg

Lane 2:

Recombinant Human octamers containing H2A at 0.5 µg

Lane 3:

Native recombinant octamers K562 cells at 3 µg

Lane 4:

Native recombinant octamers K562 cells at 1.5 µg

Lane 5:

Recombinant Human octamers containing H2A.Z.2.1 at 0.5 µg

Lane 6:

Recombinant Human octamers containing H2A.Z.1 at 0.5 µg

Secondary

All lanes:

HRP-conjugated donkey anti-rabbit IgG polyclonal at 1/10000 dilution

Predicted band size: 14 kDa

Observed band size: 15 kDa

true

Exposure time: 5min

This image is courtesy of an Abreview submitted by Ragnhild Eskeland

• WB

Lab

Western blot - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

All lanes:

Western blot - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (ab150402) at 1/5000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 15 µg

Lane 3:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates at 15 µg

Lane 4:

RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 15 µg

Lane 5:

C6 (Rat glial tumor glial cell) whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 14 kDa

Observed band size: 13 kDa

false

• WB

CiteAb

Western blot - Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (AB150402)

Histone H2A.Z western blot using anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade ab150402. Publication image and figure legend from Lowe, D. J., Herzog, M., et al., 2020, Sci Rep, PubMed 32042076.

ab150402 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab150402 please see the product overview.

Chronic γ-radiation reduces histone levels. (a) Western blot analyses of histone H2AX and γH2AX in primary fibroblasts exposed to various dose-rates of chronic γ-radiation for 7 days. Cells irradiated with a single acute dose of 4 Gy X-ray were included as control. (b) Effect of chronic γ-irradiation on H2AX levels in three different isogenic primary cell types from a different donor to that used in (a) at the same dose rates as in (a). (c) Immunoblots of other histones in chronically irradiated primary fibroblasts. (d) All significant histone level changes detected by SILAC LC-MS/MS protein analyses of samples from primary fibroblasts exposed or mock-exposed to chronic γ-radiation for 7 days.

false