MA1-010

antibody from Invitrogen Antibodies

Targeting: TRAP1 HSP75, HSP90L

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-010

Provider

Invitrogen Antibodies

Product name

TRAP1 Monoclonal Antibody (TRAP1-6)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

MA1-010 detects tumor necrosis factor receptor-associated protein (TRAP1) from human tissues. MA1-010 has been successfully used in Western blot, immunofluorescence and immunoprecipitation procedures. By Western blot, this antibody detects an ~75 kDa protein representing TRAP1. Immunofluorescence staining of TRAP1 in PC-3-M cells with MA1-010 produces a pattern consistent with mitochondrial staining. Immunoprecipitation of TRAP1 using MA1-010 fails to co-precipitate p23, Hop, or CyP40 suggesting TRAP1’s inability to associate with these co-chaperones. The MA1-010 immunogen is purified, recombinant, human TRAP1.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

TRAP1-6

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Mitochondria are devoid of poly(ADP-ribose)polymerase-1, but harbor its product oligo(ADP-ribose).

Köritzer J, Blenn C, Bürkle A, Beneke S
Journal of cellular biochemistry 2021 May;122(5):507-523

---

Cytotoxicity of withaferin A in glioblastomas involves induction of an oxidative stress-mediated heat shock response while altering Akt/mTOR and MAPK signaling pathways.

Grogan PT, Sleder KD, Samadi AK, Zhang H, Timmermann BN, Cohen MS
Investigational new drugs 2013 Jun;31(3):545-57

---

A novel C-terminal HSP90 inhibitor KU135 induces apoptosis and cell cycle arrest in melanoma cells.

Samadi AK, Zhang X, Mukerji R, Donnelly AC, Blagg BS, Cohen MS
Cancer letters 2011 Dec 22;312(2):158-67

---

Involvement of tumor necrosis factor receptor-associated protein 1 (TRAP1) in apoptosis induced by beta-hydroxyisovalerylshikonin.

Masuda Y, Shima G, Aiuchi T, Horie M, Hori K, Nakajo S, Kajimoto S, Shibayama-Imazu T, Nakaya K
The Journal of biological chemistry 2004 Oct 8;279(41):42503-15

---

Hsp90 chaperone activity requires the full-length protein and interaction among its multiple domains.

Johnson BD, Chadli A, Felts SJ, Bouhouche I, Catelli MG, Toft DO
The Journal of biological chemistry 2000 Oct 20;275(42):32499-507

---

The hsp90-related protein TRAP1 is a mitochondrial protein with distinct functional properties.

Felts SJ, Owen BA, Nguyen P, Trepel J, Donner DB, Toft DO
The Journal of biological chemistry 2000 Feb 4;275(5):3305-12

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of TRAP1 was achieved by transfecting HeLa with TRAP1 specific siRNAs (Silencer® select Product # s177). Western blot analysis (Fig. a) was performed using Whole cell extracts from the TRAP1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with TRAP1 Monoclonal Antibody (TRAP1-6) (Product # MA1-010, 1:2,000 ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000). 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 TRAP1.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-TRAP1 Monoclonal Antibody (TRAP1-6) (Product # MA1-010) and a 74kDa band corresponding to TRAP1 was observed across 4 cell lines. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), HEK-293 (Lane 3), K-562 (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2,000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of TRAP1 in NCI-H460 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a TRAP1 monoclonal antibody (Product # MA1-010) at a dilution of 1:200 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). TRAP1 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of TRAP1 in PC-3-M cells using a TRAP1 monoclonal antibody (Product # MA1-010).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

6 Figure TRAP1 interacts with oADP-ribose in mitochondria. Mitochondria isolated from 10 8 HeLaS3 cells were immunoprecipitated using TRAP1 (75 kDa) antibody. 0.5% input, 5% of final wash, 25% post-elute, and 40% of eluted material of IP either with an unrelated IgG antibody (elute control IgG) or with TRAP1 antibody (elute) were probed for oADPR presence using LP96-10. Whereas the high molecular weight signal is visible in the input fraction, this is no longer detected in the TRAP1-IP elute, but the specific signal at 75 kDa (arrow). * denotes signals from mouse IgG used for IP. IP, immunoprecipitation; PARP, poly(ADP-ribose)polymerase

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation of TRAP1 using Product # MA1-010 visualized by Coomassie Blue staining.