Anti rab7

Manufactured by Abcam

Sourced in United Kingdom

Anti-Rab7 is a primary antibody that recognizes the Rab7 protein. Rab7 is a small GTPase that regulates late endosome and lysosome function. This antibody can be used to detect and study the Rab7 protein in various experimental applications.

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Mouse anti-Rab7 (8 citations) Rabbit anti-Rab7 (5 citations) Anti-Rab7 antibody (3 citations) Mouse anti-Rab7 (Rab7-117: (2 citations)

8 protocols using anti rab7

Protein Localization and Autophagy Markers

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The following primary antibodies were used: anti-GAPDH (Sigma-Aldrich, G9545; 1:5000) anti-TOMM20 (Santa Cruz Biotechnology, SC-11415; 1:500), anti-FIS1 (Proteintech, 10956–1-1ap; 1:100), anti-ATP5F1A/ATP5A (Abcam, Ab14748; 1:200), anti-PDHA1 (Abcam, ab110330; 1:200), anti-ATG7 (Cell Signaling Technology, 8558; 1:1000), anti-RAB9A (Cell Signaling Technology, 5118; 1:1000), anti-ULK1 (Cell Signaling Technology, 8054; 1:1000), anti-RAB5A (Cell Signaling Technology, C8B1; 1:1000), anti-RAB7 (ERP7589; Abcam, ab137029; 1:1000), anti-LC3B (Sigma-Aldrich, L7543;1:200). Alexa Fluor 647‐conjugated goat anti-rabbit and anti-mouse IgG (Invitrogen, A21244 and A32728; 1:500) were used as secondary antibodies.

Godtliebsen G., Larsen K.B., Bhujabal Z., Opstad I.S., Nager M., Punnakkal A.R., Kalstad T.B., Olsen R., Lund T., Prasad D.K., Agarwal K., Myrmel T, & Birgisdottir A.B. (2023). High-resolution visualization and assessment of basal and OXPHOS-induced mitophagy in H9c2 cardiomyoblasts. Autophagy, 19(10), 2769-2788.

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Antibody Characterization for APP, BACE1, and LRP1

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The antibodies used were as follows: anti-BACE1 [AB5832, Millipore; D10E5, Cell Signaling; MAB9311, R & D systems; and NBA (Murayama et al., 2005 (link))]; anti-LRP1 1704 (Pietrzik et al., 2002 (link)); anti-APP R37 (Kametani et al., 1993 (link)); anti-rhodopsin tag 1D4 (University of British Columbia) (Farzan et al., 2000 (link); Murayama et al., 2005 (link)); anti-β-galactosidase (LacZ; MP Biomedicals); anti-β-actin (Sigma); anti-myc (Invitrogen); anti-hemagglutinin (anti-HA; rabbit: MBL; goat: Abcam); anti-flotillin-1 (IBL); anti-EEA1 (rabbit: Affinity BioReagents; goat: Biorbyt); anti-γ1-adaptin (Santa Cruz Biotechnology); anti-β-COP (Thermo Scientific); anti-rab7a (rabbit: Millipore); anti-rab7 (mouse: Abcam); and anti-GM130 (BD Biosciences).

Tanokashira D., Motoki K., Minegishi S., Hosaka A., Mamada N., Tamaoka A., Okada T., Lakshmana M.K, & Araki W. (2015). LRP1 Downregulates the Alzheimer’s β-Secretase BACE1 by Modulating Its Intraneuronal Trafficking,,. eNeuro, 2(2), ENEURO.0006-15.2015.

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Quantifying Endocytic Pathways in Cells

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Wortmannin, dynasore, SU6655, NH₄Cl, SP600125, IPA-3, MβCD and chlorpromazine were obtained from Sigma (Sigma, MO, United States), Akti-1/2 and bafilomycin A1 were obtained from Abcam (Abcam, Cambridge, United Kingdom), nystatin and EIPA was obtained from Solarbio (Solarbio, Beijing, China).
The rabbit anti-dynamin-2, anti-clathrin heavy chain, anti-Rab5, anti-Rab7, anti-PI3K p85 (phos pho Y458) + PI3 Kinase p55 (phospho Y199), anti-Akt, anti-Src, anti-Src (phospho Y416), anti-phospho-Akt (Ser473) and anti-GAPDH monoclonal antibodies were obtained from Abcam (Abcam, Cambridge, United Kingdom). The rabbit anti-JNK and anti-JNK (Thr183/Tyr185) monoclonal antibodies were obtained from Cell Signaling Technology (Cell Signaling Technology, Danvers, United States). The rabbit anti-PI3 Kinase p85 alpha polyclonal antibody, the mouse anti-Flag monoclonal antibody, goat anti-rabbit and goat anti-mouse HRP-labeled secondary antibody, Alexa fuor-488-conjugated anti-mouse, Cy™3-conjugated anti-rabbit IgG (H + L) and mounting medium with DAPI was purchased from Abcam (Abcam, Cambridge, United Kingdom). The rabbit anti-caveolin-1 monoclonal antibody was obtained from Beyotime (Beyotime Biotechnology, Shanghai, China). The mouse anti-BRSV G protein monoclonal antibody was provided by China Animal Health and Epidemiology Center.

Liu Y., Yang D., Jiang W., Chi T., Kang J., Wang Z, & Wu F. (2024). Cell entry of bovine respiratory syncytial virus through clathrin-mediated endocytosis is regulated by PI3K-Akt and Src-JNK pathways. Frontiers in Microbiology, 15, 1393127.

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Western Blot Analysis of GC Cells

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Following the instructions of the BCA protein analysis kit (Thermo Fisher Scientific, Waltham, MA, USA), GC cells were lysed in WESTERN & IP cell lysis buffer (Beyotime, Shanghai, China) and PMSF (AMRESCO, Solon, Ohio, USA). The supernatant was collected after centrifugation at 4°C for 10 min. The same amount of sample proteins was added to the wells for electrophoresis, and then transferred to the membrane by using 0.45 μm PVDF membrane (Amersham Hybond, GE Healthcare, Munich, Germany). Wells were incubated with rabbit antibody at 4°C overnight using anti-Rab7 (1-2000, Abcam, UK), PI3K (1-1000, Cell Signaling Technology, USA), pPI3K (1-1000, cell signal), AKT (1-1000, cell signal), pAKT (Ser308) (1-1000, Cell Signaling Technology, USA), and pAKT (Ser473) (1-1000, Cell Signaling Technology, USA). After washing for 10 min in TBST, the primary antibody was incubated with enhanced chemiluminescence substrate (Abcam, UK) for 2 h at room temperature, and then protein signal detection was performed using enhanced chemiluminescence (Lulong Biotech, Xiamen, China).

Liu H., Xu J., Yao Q., Zhang Z., Guo Q, & Lin J. (2020). Rab7 Is Associated with Poor Prognosis of Gastric Cancer and Promotes Proliferation, Invasion, and Migration of Gastric Cancer Cells. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 26, e922217-1-e922217-11.

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Rab GTPase Trafficking Assay

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The following primary antibodies were used; CD11a Alexa Fluor^™ 594 clone HI111 (Biolegend, San Diego, CA), rabbit polyclonal anti-Rab5, anti-Rab11A, anti-Rab7, or anti-Lamp1 antibody (Abcam, Cambridge, UK), anti-beta-actin antibody (AnaSpec, Fremont, CA) (1:1000), and anti-LtxA monoclonal antibody 107A3A3 [75 (link)] in hybridoma supernatants (1:10 dilution). The following secondary antibodies were used: goat anti-rabbit IgG Alexa Fluor^®488 (1:1000); horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Fc) or (HRP)-goat anti-rabbit (Pierce, Rockford, IL) (1:10,000). Transferrin labeled with Alexa Fluor®555 was from Invitrogen (Waltham, MA, USA). Dynamin inhibitor Dynole 34-2 and its inactive control, Dynole 31-2, were purchased from SigmaAldrich (St. Louis, MO), Dynasore and Pitstop 2 (Abcam, Cambridge, UK). The inhibitors were used in the following concentrations: 10 μM Dynole 34-2; 10 μM Dynole 31-2; 10 μM Dynasore; 5 μM Pitstop 2.

Lally E.T., Boesze-Battaglia K., Dhingra A., Gomez N.M., Lora J., Mitchell C.H., Giannakakis A., Fahim S.A., Benz R, & Balashova N. (2020). Aggregatibacter actinomycetemcomitans LtxA Hijacks Endocytic Trafficking Pathways in Human Lymphocytes. Pathogens, 9(2), 74.

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Osteoclast Immunostaining Protocols

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BMMs were cultured on glass or dentine slides and induced to differentiate into osteoclasts. Osteoclasts were fixed with 4% paraformaldehyde for 15 min then washed three times with phosphate buffered saline with Tween 20 (PBST). Cells were permeabilized in 0.1% Triton-X for 10 min and blocked for 1 h in PBST containing 2.5% bovine serum albumin (BSA) and 10% normal goat serum. Primary antibody diluted in Can Get Signal immunostain solution A (Toyobo) was added for 12 h at 4 °C, followed by 1 h incubation with secondary antibody together with phalloidin-647 (Abcam) and DAPI. The following antibodies were used: anti-coronin 1A (Abcam), anti-cathepsin K (Abcam), anti-LAMP1 (Cell Signaling Technology, Biolegend), anti-LC3 (MBL), anti-rab7 (Abcam), anti-rab27a (R&D), anti-mouse Alexa Fluor 488 and anti-rabbit Alexa Fluor 594 (both from Molecular Probes). Slides were imaged on LSM 710 (Zeiss) and analysed using MetaMorph software.

Ohmae S., Noma N., Toyomoto M., Shinohara M., Takeiri M., Fuji H., Takemoto K., Iwaisako K., Fujita T., Takeda N., Kawatani M., Aoyama M., Hagiwara M., Ishihama Y, & Asagiri M. (2017). Actin-binding protein coronin 1A controls osteoclastic bone resorption by regulating lysosomal secretion of cathepsin K. Scientific Reports, 7, 41710.

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Phagocytosis Analysis with Antibodies

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PhagoFACS assays: (additional antibodies) rabbit polyclonal anti-ovalbumin (OVA) (Sigma Aldrich); mouse monoclonal anti-Rab7 (Abcam); rabbit polyclonal anti-BSA (Invitrogen).

Weimershaus M., Mauvais F.X., Evnouchidou I., Lawand M., Saveanu L, & van Endert P. (2020). IRAP Endosomes Control Phagosomal Maturation in Dendritic Cells. Frontiers in Cell and Developmental Biology, 8, 585713.

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Detecting HIV-1 and Protein Knockdown

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HIV-1 p24 was detected using anti-HIV-1 core antigen antibody-FITC (KC57-FITC; Beckman Coulter), and actin labeled with Cytopainter Phallodin-iFluor-555 (Abcam). Protein knockdown was detected by immunoblotting using rabbit anti-ARF1, anti-BIN1, anti-RAB8A, mouse anti-Rab7L1 (Abcam), and actin (Merck) antibodies, followed by secondary HRP-conjugated goat anti-rabbit and anti-mouse antibodies (Dako). Confocal microscopy was carried out using the primary antibodies anti-human CD81-APC (BD), anti-EEA1, anti-CHMP2B, anti-LAMP1, anti-Rab7, anti-Rab11, anti-Rab5 (Abcam). HRP uptake was detected using anti-HRP (Jackson Immunolaboratory). All unlabeled primary antibodies were detected with secondary anti-rabbit Alexa Fluor 546 (Life Technologies). The pharmacological inhibitors LY294002, bafilomycin A1, and indinivir (Sigma-Aldrich) were used at 50 μM, 0.5 μM, and 2 μg/ml, respectively.

Bayliss R., Wheeldon J., Caucheteux S.M., Niessen C.M, & Piguet V. (2020). Identification of Host Trafficking Genes Required for HIV-1 Virological Synapse Formation in Dendritic Cells. Journal of Virology, 94(9), e01597-19.

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