71-5300

antibody from Invitrogen Antibodies

Targeting: RAB11A YL8

Provider product page for 71-5300

Western blot

ELISA

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Flow cytometry

Immunoelectron microscopy

Other assay

Antibody data

Antibody Data
Antigen structure
References [95]
Comments [0]
Validations
Immunocytochemistry [5]
Flow cytometry [1]
Other assay [3]

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Product number: 71-5300 - Provider product page
Provider: Invitrogen Antibodies
Product name: RAB11A Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: This polyclonal antibody reacts specifically with the human Rab11A protein. Cross-reactivity with other Rab family members has not been detected. Positive control: Human HEK-293 cells
Reactivity: Human, Mouse, Rat, Canine, Rabbit
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 0.25 mg/mL
Storage: -20°C

RAB11A protein structure - 71-5300 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Rab11 (green) in 3T3 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes and blocked with 3% Blocker BSA (Product # 37525) in PBS for 30 minutes at room temperature. Cells were stained with or without Rab11 rabbit polyclonal antibody (Product # 71-5300), at a concentration of 5 µg/mL for 1 hour at room temperature, and then incubated with a Goat anti-Rabbit (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:1000 for 1 hour at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Ras-related protein Rab-11A was performed using 70% confluent log phase Hep G2 cells. 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 RAB11A Polyclonal Antibody (Product # 71-5300) at 1:100 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), (1:2,000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic (Golgi complex and cleavage furrow like pattern) localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7A1110LZR) and externally deconvoluted (D.Sage et al. / Methods 115 (2017) 28–41).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Ras-related protein Rab-11A was performed using 70% confluent log phase U-87 MG cells. 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 RAB11A Polyclonal Antibody (Product # 71-5300) at 1:100 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), (1:2,000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7A1110LZR) and externally deconvoluted (D.Sage et al. / Methods 115 (2017) 28–41).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Ras-related protein Rab-11A was performed using 70% confluent log phase A-431 cells. 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 RAB11A Polyclonal Antibody (Product # 71-5300) at 1:100 in 0.1% BSA, incubated at 4 ° C overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7A1110LZR) and externally deconvoluted (D.Sage et al. / Methods 115 (2017) 28-41).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Rab11 was done on 70% confluent log phase U2OS cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Rab11 Rabbit Polyclonal Antibody (Product # 71-5300) at 2 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6. Downregulation of calsyntenin-1 by RNAi results in enhanced APP processing and increased secretion of Abeta. Cortical neurons were infected with rAAVs expressing a hairpin targeting calsyntenin-1 (shCst1) or a nonsense hairpin (shNS), together with eGFP. ( A ) Western blot of cell lysates. The signal indicated equal expression of the recombinant proteins. ( B ) Quantification of Western blots. Values indicate mean+-s.e.m. (*, P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 9 Characterization of purified ICVs. A549 cells were infected with the WSN virus and submitted to subcellular fractionation at 14 hpi. a Western-blot analysis of subcellular fractions of mock-infected (left panel) and IAV-infected (right panel) A549 cells, upon sucrose gradient ultracentrifugation. The same amount of total protein was analyzed for each fraction. TfR: transferrin receptor; PDI: protein disulfide isomerase; Cal: calreticulin; Clathrin HC: Clathrin Heavy Chain. For gel source data, Supplementary Fig. 17 . b TEM analysis of an aliquot of fraction 60-40% from IAV-infected cells, upon fixation and negative staining. Black arrows: ICVs. White arrows: vRNPs. Scale bar: large panel 200 nm, small pannel 100 nm

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1. Calsyntenin-1/APP vesicles contain BACE1, but no ADAM10. ( A ) Schematic representation of vesicle immunoisolation. Vesicle immunoisolations were performed with antibodies against cytosolic epitopes of vesicle-associated proteins and Dynabeads coated with protein A. Captured vesicles were isolated and washed. Vesicular proteins were eluted by lysis of the captured vesicles with a mild detergent. Note that the vesicular component bearing the antigen targeted for immunoisolation is not or only partially found in the eluate, because of its strong binding to the antibody/protein A/bead complex. Its complete elution required harsher conditions, e.g. boiling of the beads in SDS. Therefore, the antigen used for vesicle immunoisolation is not shown on the Western blots of the (mild) eluates. Examples of antigen elutions under harsh condition from the beads are shown in the panels labeled ""beads"". ( B ) Specificity tests of anti-APP, anti-ADAM10, and anti-BACE1 antibodies, performed by the addition of ~25 ug of recombinant antigen (GST-APPcyto, GST-ADAM10cyto, BACE1[485-501]). Saturation with recombinant antigen prevented vesicle capture. ( C ) Calsyntenin-1 organelles contained BACE1, but no ADAM10 (black box). ( D ) APP and syntaxin13 immunoisolates contain a low level of active ADAM10 and abundant BACE1. Neither BACE1 nor ADAM10 were found in Rab11 immunoisolates. ( E ) Rab5 immunoisolates contain a small amount of active ADAM10 (arrowhead) and abundant BACE1 (green box).