- PA5-29022 - Invitrogen Antibodies RAB5A antibody

Submitted references RAB5A expression is a predictive biomarker for trastuzumab emtansine in breast cancer.

Engebraaten O, Yau C, Berg K, Borgen E, Garred Ø, Berstad MEB, Fremstedal ASV, DeMichele A, Veer LV', Esserman L, Weyergang A
Nature communications 2021 Nov 5;12(1):6427

Trappc9 deficiency in mice impairs learning and memory by causing imbalance of dopamine D1 and D2 neurons.

Ke Y, Weng M, Chhetri G, Usman M, Li Y, Yu Q, Ding Y, Wang Z, Wang X, Sultana P, DiFiglia M, Li X
Science advances 2020 Nov;6(47)

Cancer Alters the Metabolic Fingerprint of Extracellular Vesicles.

Palviainen M, Laukkanen K, Tavukcuoglu Z, Velagapudi V, Kärkkäinen O, Hanhineva K, Auriola S, Ranki A, Siljander P
Cancers 2020 Nov 6;12(11)

Cutaneous T-Cell Lymphoma (CTCL) Cell Line-Derived Extracellular Vesicles Contain HERV-W-Encoded Fusogenic Syncytin-1.

Laukkanen K, Saarinen M, Mallet F, Aatonen M, Hau A, Ranki A
The Journal of investigative dermatology 2020 Jul;140(7):1466-1469.e4

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of RAB5A using 30 µg of A) A431 and B) Raji lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a RAB5A polyclonal antibody (Product # PA5-29022) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on membrane enriched cell extracts (30 µg lysate) of MCF-7 (Lane 1), HeLa (Lane 2), HEK293 (Lane 3), Jurkat (Lane 4), SK-BR-3 (Lane 5), MDA-MB-231 (Lane 6), THP-1 (Lane 7) and tissue extract of Rat Brain (Lane 8). The blot was probed with Anti-RAB5 Polyclonal Antibody (Product # PA5-29022, 1:2000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 25 kDa band corresponding to RAB5 was observed across all the cell lines and tissue tested. A non-specific band was observed at 50 kDa for all cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using RAB5 Polyclonal Antibody (Product # PA5-29022). Sample (50 µg of whole cell lysate). Lane A: mouse brain. 12% SDS PAGE. RAB5 Polyclonal Antibody (Product # PA5-29022) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using RAB5 Polyclonal Antibody (Product # PA5-29022). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with RAB5 Polyclonal Antibody (Product # PA5-29022) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of RAB5 was performed by separating 30 µg of various whole cell lysates by 12 % SDS-PAGE. Proteins were transferred to a membrane and probed with a RAB5 Polyclonal Antibody (Product # PA5-29022) at a dilution of 1:1000. A. 293T, B. A431 , C. HeLa , D. A375, E. NCI-H929.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using RAB5 Polyclonal Antibody (Product # PA5-29022). Jurkat whole cell extracts and Jurkat exosome extract (15.4 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with RAB5 Polyclonal Antibody (Product # PA5-29022) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of RAB5 was performed by separating 30 µg of various whole cell lysates by 12 % SDS-PAGE. Proteins were transferred to a membrane and probed with a RAB5 Polyclonal Antibody (Product # PA5-29022) at a dilution of 1:1000. A. Neuro2A, B. BCL-1, C. Raw264.7, D. C2C12.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of RAB5 was performed by separating 30 µg of whole cell lysates by 12 % SDS-PAGE. Proteins were transferred to a membrane and probed with a RAB5 Polyclonal Antibody (Product # PA5-29022) at a dilution of 1:1000. A. PC-12, B. Rat-2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of RAB5 was achieved by transfecting MCF cells with RAB5 specific siRNAs (Silencer® select Product # s11680). Western blot analysis (Fig. a) was performed using whole cell extracts from the RAB5 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with RAB5 Polyclonal Antibody (Product # PA5-29022, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/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 RAB5. [Note: Non-specific band observed at ~50Kda].

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: MRC-5 cells (fetal lung fibroblasts) were grown on glass coverslips in 6 well dishes and infected for 48 h with VZV. Cells were fixed in 2% Paraformaldehyde and permeabilized with 0.02% Triton X-100 for 1 h at room temperature. Blocked cells for 2 h at RT in 5% non-fat milk plus 2.5% normal goat serum in PBS. Anti-VZV gE mouse monoclonal antibody (red) and anti-Rab5 rabbit polyclonal antibody (green, Product # PA5-29022, 1:100 in PBS) were incubated 2 h at RT (with rocking), then overnight at 4oC (with gentle rocking). After washing with PBS, cells were incubated with goat anti rabbit AlexaFluor488 (1:1250), goat anti mouse AlexaFluor546 (1:1250) and Hoechst 33342 (blue, 1:1000) for 2 h at RT (rocking, in the dark). Washed the cells with PBS. Mounted coverslips to glass and viewed on Zeiss 710 Laser Scanning Confocal Microscope. Images shown are at 400X (upper) or 630X (bottom) total magnification, respectively. Data courtesy of Dr. Grose’s lab.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: RAB5 Polyclonal Antibody detects RAB5A protein at cytosol on U87 xenograft by immunohistochemical analysis. Sample: Paraffin-embedded U87 xenograft. RAB5 Polyclonal Antibody (Product # PA5-29022) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Rab5A antibody immunoprecipitates Rab5A protein in IP experiments. IP Sample: 1 mg HeLa whole cell lysate/extract A. 30 µg HeLa whole cell lysate/extract B. Control with 2 µg of preimmune rabbit IgG C. Immunoprecipitation of Rab5A protein by 2 µg of Rab5A antibody (Product # PA5-29022) 15% SDS-PAGE The immunoprecipitated Rab5A protein was detected by Rab5A antibody (Product # PA5-29022) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 2 HER2 and RAB GTPase expression in cell lines. A Representative Western blot of HER2 and gamma-tubulin expression in SK-BR-3, SKOV-3, HCC1954, AU-565, MDA-MB-453, and MDA-MB-231 cells ( n = 3). B Quantification of the HER2 Western blots relative to those of gamma-tubulin. Data points represent the values of three independent experiments, the bars represent the average and the error bars represent the SD of the mean. C Linear regression analysis curve of HER2 protein expression and T-DM1 sensitivity (1/IC 50 (T-DM1)). D Representative western blot ( n = 3) of RAB4A, RAB5A, RAB11A, and gamma-tubulin expression in SK-BR-3, SKOV-3, AU-565, HCC1954, and MDA-MB-453 cells. E - G Quantification of the RAB4, RAB5, and RAB11 Western blots relative to those of gamma-tubulin. Data points represent the values of three independent experiments, the bars represent the average and the error bars represent the SD of the mean. Source data are provided as a Source Data file.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Characterization of EVs. ( A ) Nanoparticle tracking analysis of total amount and size distribution of EVs. Four replicates of each EV type were analyzed. ( B ) Western blot analysis of TSG101 and RAB5 (conventional EV markers) revealed that these markers were present in all EV samples, while the mitochondrial protein TOM20 was present only in the cell lysate used as a control. Total of 1.0E10 EVs were loaded in each well. ( C ) Immunoelectron microscopy revealed the presence of the EV marker CD63 on the surface of EVs by 10-nm colloidal gold particles (red arrowheads). Immunostaining without the primary antibody was used as a negative control for CD63 immunoelectron microscopy.

PA5-29022

Antibody data