MA1-2022

antibody from Invitrogen Antibodies

Targeting: H2AX H2AFX

Provider product page for MA1-2022

Western blot

ELISA

Immunocytochemistry

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [20]
Comments [0]
Validations
Immunocytochemistry [8]
Immunohistochemistry [1]
Other assay [9]

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Product number: MA1-2022 - Provider product page
Provider: Invitrogen Antibodies
Product name: Phospho-Histone H2A.X (Ser139) Monoclonal Antibody (3F2)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: MA1-2022 was used in IHC to successfully detect H2A.X pSer140 in postnatal mouse lung section. MA1-2022 detects human and mouse phosphorylated H2AX. MA1-2022 has been used successfully in Western blot, immunofluorescence, and ELISA procedures. By Western blot this antibody detects ~17 kDa protein representing phosphorylated H2AX in gamma irradiated HeLa cell lysate. In immunofluorescence procedures, MA1-2022 recognizes phosphorylated H2AX in gamma irradiated HeLa cells. ELISA of phosphorylated H2AX can also be performed using MA1-2022. MA1-2022 was used in IHC to successfully detect H2A.X pSer140 in postnatal mouse lung section. The MA1-2022 immunogen is a synthetic peptide sequence surrounding phosphorylated Ser140. Some references call Ser140, by the name Ser139. This antibody was orginally validated as part of a Thermo Scientific Cellomics High Content Screening Kit. The antibody sold separately may have slightly different performance and may need to be further optimized for the best results.
Reactivity: Human, Mouse
Host: Mouse
Isotype: IgG
Antibody clone number: 3F2
Vial size: 100 μg
Concentration: 1 mg/mL
Storage: -20°C, Avoid Freeze/Thaw Cycles

H2AX protein structure - MA1-2022 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Phospho-Histone H2A.X (Ser140) using a monoclonal antibody (Product # MA1-2022).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence staining of Phospho-H2AX in A549 cells. Cells were treated with vehicle (control; 0.1% DMSO in media) or with 50 µM etoposide for 1 hour

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence staining using Product # MA1-2022, treatment with paraquat and iron induces MnSOD and Phosphorylation of H2AX in RAW 264.7 macrophages. Cells were treated for 20 hours with paraquat (500 µM) and iron (200 µg/mL) and stained with anti-Phospho-H2AX antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence image of phosphorylated H2AX in gamma irradiated HeLa cells using Product # MA1-2022.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence image of phosphorylated H2AX in gamma irradiated HeLa cells using Product # MA1-2022.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence staining of Phospho-H2AX in A549 cells. Cells were treated with vehicle (control; 0.1% DMSO in media) or with 50 µM etoposide for 1 hour

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence staining using Product # MA1-2022, treatment with paraquat and iron induces MnSOD and Phosphorylation of H2AX in RAW 264.7 macrophages. Cells were treated for 20 hours with paraquat (500 µM) and iron (200 µg/mL) and stained with anti-Phospho-H2AX antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Phospho-Histone H2A.X (Ser140) using a monoclonal antibody (Product # MA1-2022).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5. Int-C3CNLS does not induce DNA damage or cytotoxicity. ( A ) MTT-based cell proliferation assays were performed to assess effects on cell proliferation rates upon Int expression in human cell lines. HT1080 cells untransfected (HT1080), and FACS sorted GFP + cells obtained after co-transfection of pCMV-EGFP with either pCMVssInt-Ina (INA; expressing inactive integrase) or pCMVssInt-C3CNLS (C3CNLS) were analyzed for the effect on cellular proliferation using the colorimetric MTT assays over the indicated time course. Data show the mean of triplicates and standard deviation of a representative experiment. n = 2. ( B ) Western blot analysis to determine phospho-gamma-H2AX levels to assess DNA damage induced by expression of Int in A549 cells. Cell lysates prepared at time points of 24, 48 and 72 h (post transfection) from A549 cells transfected with pCMVssInt-Ina or pCMVssInt-C3C (expressing Int-C3CNLS) and from control cells treated with the carrier (Lipofectamine2000 Transfection reagent) were subjected to western blot analysis using antibodies against phospho-gamma-H2AX (top panel). UT, untreated cells as negative control; HU, cells treated with hydroxy urea (10 mM for 24 h) as positive control; M, Marker lane. beta-actin was used as loading control (bottom panel). ( C ) Western blot analysis to determine phospho-gamma-H2AX levels to assess DNA damage induced by expression of Int in HT1080 cells. Forty-eight hours post transfection, top and 72 h post transfection, bo

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 Induction of active caspase-3 and/or gammaH2AX by OA, DTX-1 and -2 in Caco-2 and HT29-MTX cells. Representative images at 10x magnification of Caco-2 and HT29-MTX cells treated 24 h with 150 nM OA ( A ); DAPI staining in blue corresponds to the nucleus. Caco-2 and HT29-MTX cells were treated with increasing concentrations of OA, DTX-1 and -2 for 24 h and were then labeled for detection of active caspase-3 (green) and gammaH2AX (red) by high content analysis ( B ). The percentage of positive cells for active caspase-3 and gammaH2AX was determined using a threshold based on control conditions. Data are expressed as stacked bars representing unlabeled cells (white), active caspase-3-positive cells (green), gammaH2AX-positive (red) and positive for both active caspase-3 and gammaH2AX (orange). n = 3.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: (A) The Western blot analysis demonstrates the rescue of gamma-irradiation-induced expression and phosphorylation of ATM, the phosphorylation of H2AX, the expression of p53, Noxa and E2F1 as well as the phosphorylation of CHK1 and 2 in response to the induction of HURP expression by the addition of Dox to the culture media of LNCaP-TetOn-HURP cells. Actin was used internal control for loading and transfer. EMSA demonstrates the inhibition of gamma-irradiation-induced DNA binding activities of the transcription factors E2F1 (B), p53 (C,D) EMSA demonstrates the rescue of gamma-irradiation-induced DNA- binding activity of the transcription factor p53 in response to the induction of HURP expression by the addition of Doxycycline to the culture media of DU145-TetOn-HURP cells. (E) EMSA analysis demonstrated the ability of HURP to inhibit gamma-irradiation -induced DNA binding activity of the transcription factor p53 in LNCap and DU145 cells, whereas in p53- PC3 cells is not detectable compared to positive control. The data are representative of three independent experiments. (F,G) Immune fluorescence demonstrates the inhibition of gamma-irradiation-induced H2AX (red) in response to the induction of HURP expression (green) by the addition of Dox to the culture media of LNCaP-TetOn-HURP cells. The nucleus is stained with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 DNA damage induced by X-ray radiation. ( A ) Bright field image depicting the uptake of different concentrations of NPs ( B - E ) A375 cells treated with different concentrations of NPs and irradiated with 0, 2, 4, 6 Gy and stained for gamma-H2AX. ( F ) Quantification of number of foci/cell using ImageJ software. ( G ) Clonogenic assay of samples treated with 0, 25, 50, 100 mug/mL Y 2 O 3 and exposed to 0, 2, 4, 6 Gy doses of radiation (* p < 0.05, *** p < 0.005, **** p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 a Example of dermal sample after plasma treatment, stained for gammaH2AX. Positive nuclei are indicated (arrow head); bar is 50 mum. b Quantification of the relative number of cells (%) positive for gammaH2AX after plasma treatment of dermal tissue ( n =4-5 in duplicate). c Re-epithelialization and d proliferation of keratinocytes in BWM after 2 weeks of culture. Plasma was applied four times in 10 days. Mean values and standard deviation are shown ( n =6 for 0 and 120 s, n =4 for 60 s, all in duplicate). Significant differences were not observed

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 Effect of crude extract of V. rugosum , the aqueous methanol fraction and PnTX-G on ( a ) Ki-67 expression and on ( b ) gammaH2AX phosphorylation in Caco-2 cells. Data are presented as the mean average nuclear intensity of Ki-67 and gammaH2AX immunofluorescence +- SEM.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Effect of purified fractions of V. rugosum on Ki-67 expression ( a ) and on gammaH2AX phosphorylation ( b ) in Caco-2 cells. Data are presented as the mean average nuclear intensity of Ki-67 and gammaH2AX immunofluorescence +- SEM.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 Representative images of ( a ) gammaH2AX immunofluorescence in untreated Caco-2 cells; ( b ) Caco-2 cells treated with F3 at 156 ng mL -1 ; ( c ) Ki-67 immunofluorescence in untreated Caco-2 cells; ( d ) Caco-2 cells treated with F3 at 156 ng mL -1 .