Anti-Rab5 抗体 [EPR5438] - Early Endosome Marker

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (AB109534)

Immunohistochemical staining of paraffin embedded human pancreas with purified ab109534 at a working dilution of 1 in 1000. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• WB

Unknown

Western blot - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (AB109534)

All lanes:

Western blot - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (ab109534) at 1/1000 dilution

Lane 1:

293T cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Predicted band size: 24 kDa

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• WB

Lab

Western blot - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (AB109534)

ab109534 was shown to react with RAB5A in wild-type HAP1 cells in Western blot with loss of signal observed in a RAB5A knockout cell line. Wild-type HAP1 and RAB5A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab109534 overnight at 4 °C at a 1000 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (ab109534) at 1/1000 dilution

Lane 1:

Wild-type HAP1 lysate at 20 µg

Lane 2:

RAB5A Knockout HAP1 lysate at 20 µg

Observed band size: 24 kDa

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• WB

Lab

Western blot - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (AB109534)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (ab109534) at 1/5000 dilution

Lane 1:

Mouse brain at 10 µg

Lane 2:

Rat brain at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 24 kDa

Observed band size: 24 kDa

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• IHC-P

AbReview57502****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (AB109534)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Pancreas tissue sections (with ADM and PanIN lesions) from KC mice labeling Rab5 with ab109534 at 1/200 dilution. Tissue was fixed with paraformaldehyde and permeabilized with TritonX100 0.1% for 10 min at room temperature. Heat-mediated antigen retrieval was performed using TRIS-EDTA pH9 buffer. Tissue sections were blocked with 10% goat serum for 1 hour at 20°C. A polyclonal goat anti-rabbit Alexa Fluor 488 secondary antibody was used at 1/500 dilution.

This image is courtesy of an anonymous abreview.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (AB109534)

Immunohistochemical staining of Rab5 in paraffin embedded human brain tissue with unpurified ab109534, at 1/100 dilution.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (AB109534)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (ab109534) at 1/5000 dilution

Lane 1:

MCF7 cell lysate at 10 µg

Lane 2:

HEK293 cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (AB109534)

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : Rab5 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : Hu brain whole cell lysate (20 μg)
Lane 4 : Ms brain whole cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab109534 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

All lanes:

Western blot - Anti-Rab5 antibody [EPR5438] - Early Endosome Marker (ab109534)

Predicted band size: 24 kDa

false