PA5-87938

antibody from Invitrogen Antibodies

Targeting: MTAP c86fus, MSAP

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-87938

Provider

Invitrogen Antibodies

Product name

MTAP Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: MASGTTTTAV KIGIIGGTGL DDPEILEGRT EKYVDTPFGK PSDALILGKI KNVDCVLLAR HGRQHTIMPS KVNYQANIWA LKEEGCTHVI VTTACGSLRE EIQPGDIVII DQFIDRTTMR PQSFYDGSHS CARGVCHIPM AEPFCPKTRE VLIETAKKLG LRCHSKGTMV TIEGPRFSSR AESFMFRTWG ADVINMTTVP EVVLAKEAGI CYASIAMATD YDCWKEHEEA VSVDRVLKTL KENANKAKSL LLTTIPQIGS TEWSETLHNL KNMAQFSVLL PRH; Positive Samples: 22Rv1, 293T, Mouse liver; Cellular Location: Cytoplasm, Nucleus

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.48 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Proteomics Profiling of KAIMRC1 in Comparison to MDA-MB231 and MCF-7.

Alghanem B, Ali R, Nehdi A, Al Zahrani H, Altolayyan A, Shaibah H, Baz O, Alhallaj A, Moresco JJ, Diedrich JK, Yates JR 3rd, Boudjelal M
International journal of molecular sciences 2020 Jun 18;21(12)

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of MTAP in extracts of various cell lines using MTAP Polyclonal Antibody (Product # PA5-87938) at a dilution of 1:1000. A HRP Goat Anti-Rabbit IgG (H+L) secondary antibody was used at a dilution of 1:10,000. Lysates/proteins: 25 µg per lane. Blocking buffer: 3% nonfat dry milk in TBST.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-MTAP Polyclonal Antibody (Product # PA5-87938) and a 32 kDa band corresponding to S-methyl-5-thioadenosine phosphorylase was observed across all positive cell lines tested, and not in negative cell lines (MCF7 and Jurkat). Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), MCF7 (Lane 3), Jurkat (Lane 4), HEK-293 (Lane 5), HepG2 (Lane 6), Raji (Lane 7) and NIH 3T3 (Lane 8) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry-Immunofluorescence analysis of MTAP was performed in A-549 cells using MTAP Polyclonal Antibody (Product # PA5-87938).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of S-methyl-5-thioadenosine phosphorylase was performed using 70% confluent log phase HeLa and MCF7 cell lines. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with MTAP Polyclonal Antibody (Product # PA5-87938) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), 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 dilution). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e represents MCF7 cell line which does not express MTAP protein. 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 analysis of MTAP in paraffin-embedded rat brain using MTAP Polyclonal Antibody (Product # PA5-87938) at a dilution of 1:100.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of MTAP in paraffin-embedded mouse liver using MTAP Polyclonal Antibody (Product # PA5-87938) at a dilution of 1:200.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5 Expression of MTAP, PARP1, KCTD12, and Lamin A/C in cancer cell lines: Western blot analysis of protein expression of MTAP, PARP1, KCTD12, Lamin A/C, and phosphorylated Lamin A/C in MDA MB231, MCF-7, and KAIMRC1 cells. Preparation of cell lysates and western blot were performed as described under the Materials and Methods section. In both normal (+) and serum-starved (-) conditions, KAIMRC1 cells showed strong expression of MTAP, PARP1, and KCTD12 compared to MDA-MB231 and MCF-7 cells. Moreover, KAIMRC1 cells showed weak expression of Lamin A/C and phosphorylated Lamin A/C in comparison to MDA MB231 cells, thus validating our proteomics results.