MA1-23261

antibody from Invitrogen Antibodies

Targeting: THOC1 HPR1, P84

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-23261

Provider

Invitrogen Antibodies

Product name

Nuclear Matrix Protein p84 Monoclonal Antibody (5E10)

Antibody type

Monoclonal

Antigen

Other

Description

Suggested positive controls are HeLa, Raji, and MOLT4.

Reactivity

Human, Mouse, Rat, Hamster

Host

Mouse

Isotype

IgG

Antibody clone number

5E10

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Dihydroartemisinin inhibits the viability of cervical cancer cells by upregulating caveolin 1 and mitochondrial carrier homolog 2: Involvement of p53 activation and NAD(P)H:quinone oxidoreductase 1 downregulation.

Zhang T, Hu Y, Wang T, Cai P
International journal of molecular medicine 2017 Jul;40(1):21-30

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of p84 using 30 µg of A) Jurkat (B) Raji (C) 293T (D) A431 (E) HeLa (F) HepG2 (G) H1299 (H) HCT116 (I) MCF-7 (J) NT2D1 (K) PC-3 and L) U87-MG lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a p84 monoclonal antibody (Product # MA1-23261) at a dilution of 1:1000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of p84 using 30 µg of A) 293T (B) NIH-3T3 (C) mouse brain and D) PC-12 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a p84 monoclonal antibody (Product # MA1-23261) at a dilution of 1:1000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of p84 using 30 µg of cell lysate of A) HeLa (B) HeLa cytosol fraction and C) HeLa nucleus fraction lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a p84 monoclonal antibody (Product # MA1-23261) at a dilution of 1:1000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of p84 using A) 30 µg 293T whole cell lysate (B) 30 µg A431 whole cell lysate (C) 30 µg HeLa whole cell lysate (D) 30 µg HepG2 whole cell lysate and E) 30 µg A375 whole cell lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a p84 monoclonal antibody (Product # MA1-23261) at a dilution of 1:500.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of K-562 (Lane 1), Jurkat (Lane 2), A-431 (Lane 3), Hep G2 (Lane 4), A-375 (Lane 5), HeLa (Lane 6), NIH/3T3 (Lane 7), HCT 116 (Lane 8), MCF7 (Lane 9) and HEK-293 (Lane 10). The blot was probed with Anti-Nuclear Matrix Protein p84 Monoclonal Antibody (Product # MA1-23261, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A 75 kDa band corresponding to Nuclear Matrix Protein p84 was observed across the cell lines tested.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261). Sample (30 µg of whole cell lysate). Lane A: HeLa. Lane B: HeLa nucleus. 7.5% SDS PAGE. Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261) diluted at 1:1,000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261). Sample (50 µg of whole cell lysate). Lane A: mouse Cerebellum. 7.5% SDS PAGE. Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261) diluted at 1:1,000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Detection of human p84/N5 in nuclear fraction by anti-p84/N5 5E10 monoclonal antibody Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261) in western blot experiment. Lane 1: total lysate, Lane 2: cytoplasmic fraction, Lane 3: membrane fraction, Lane 4: nuclear fraction.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261). Sample (whole cell lysate). Lane A: 293T 20 µg. Lane B: 293T 10 µg. Lane C: 293T 5 µg. 7.5% SDS PAGE. Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261) diluted at 1:1,000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of Nuclear Matrix Protein p84 was achieved by transfecting HEK-293 cells with Nuclear Matrix Protein p84 specific siRNAs (Silencer® select Product # s19396; s19397). Western blot analysis (Fig. a) was performed using whole cell extracts from the Nuclear Matrix Protein p84 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Nuclear Matrix Protein p84.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry-Immunofluorescence analysis of Nuclear Matrix Protein p84 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1 23261) diluted at 1:500. Red: phalloidin, a cytoskeleton marker.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Nuclear Matrix Protein p84 was performed using 70% confluent log phase HCT 116 cells treated with 0.5 µg of Brefeldin for 24 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Nuclear Matrix Protein p84 Mouse Monoclonal Antibody (Product # MA1-23261) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nuclear localization. Panel e shows untreated cells with less Nuclear signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry (Paraffin) analysis of Nuclear Matrix Protein p84 was performed in paraffin-embedded human breast carcinoma tissue using Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261) at a dilution of 1:200. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry (Paraffin) analysis of Nuclear Matrix Protein p84 was performed in paraffin-embedded human lung cancer tissue using Nuclear Matrix Protein p84 Monoclonal Antibody (5E10) (Product # MA1-23261) at a dilution of 1:100. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

p84 antibody [5E10] immunoprecipitates p84 protein in IP experiments. IP Sample: HepG2 whole cell lysate/extract A : 30 µg whole cell lysate/extract of p84 protein expressing HepG2 cells B : Control with 3 µg of pre-immune mouse IgG C : Immunoprecipitation of p84 by 3 µg of p84 antibody [5E10] (Product # MA1-23261) 7.5% SDS-PAGE The immunoprecipitated p84 protein was detected by p84 antibody [5E10] (Product # MA1-23261) diluted at 1 : 1,000.