Anti-RAB27A + RAB27B antibody [1G7]

• WB

Lab

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (AB55667)

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : RAB27A knockout HAP1 whole cell lysate (20 μg)
Lane 3 : Jurkat whole cell lysate (20 μg)
Lane 4 : MCF7 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab55667 observed at 27 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab55667 was shown to specifically react with RAB27A when RAB27A knockout samples were used. Wild-type and RAB27A knockout samples were subjected to SDS-PAGE. ab55667 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 2.5 ug/ml and 1/10000 dilution respectively. Blots were developed with IRDye^® 800CW Goat anti-Mouse IgG (H + L) and IRDye^® 680 Goat anti-Rabbit IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (ab55667)

Predicted band size: 25 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB27A + RAB27B antibody [1G7] (AB55667)

Anti-RAB27A + RAB27B antibody [1G7] (ab55667) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human lymphoma. Heat induced antigen retrieval with Citrate buffer pH 6 has been performed prior to immunostaining.

This image was generated using the ascites version of the product.

• WB

Supplier Data

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (AB55667)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (ab55667)

All lanes:

HL-60 cell lysate

Predicted band size: 25 kDa

false

• WB

Lab

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (AB55667)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Anti-Flag tag antibody was used as a loading control at a 1/100,000 dilution. Anti-6X His tag® antibody [EPR20547] - ChIP Grade, ab213204 was used at 1/5,000 dilution.

All lanes:

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (ab55667) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector contain a His-tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human RAB27A expression vector containi a His-tag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human RAB27B expression vector containi a Flag-tag whole cell lysate at 20 µg

Secondary

Lanes 1 - 3:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Lanes 1 - 3:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/20000 dilution

Observed band size: 27 kDa

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (AB55667)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (ab55667)

Lane 1:

RAB27A transfected lysate(24.9 KDa)

Lane 2:

Non-transfected lysate

Predicted band size: 25 kDa,41 kDa

Observed band size: 38 kDa

false

• WB

Supplier Data

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (AB55667)

GAPDH (36.1 kDa) used as specificity and loading control.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (ab55667)

Lane 1:

RAB27A over-expressed 293 cell line, cotransfected with RAB27A Validated Chimera RNAi

Lane 2:

Non-transfected control

Predicted band size: 25 kDa

false

• WB

AbReview9941****

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (AB55667)

Rab27a (25kDa)+GFP(27kDa)=52kDa

The blot was blocked with 5% milk for 30 minutes at 25°C and incubated with the primary antibody for 12 hours at 4°C.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (ab55667) at 1/200 dilution

Lane 1:

whole cell lysate of HEK 293 transfected with empty pCDNA3 vector at 20 µg

Lane 2:

whole cell lysate of HEK 293 cells expressing Rab27a-GFP at 20 µg

Secondary

All lanes:

HRP conjugated goat anti-mouse

Predicted band size: 25 kDa

Observed band size: 52 kDa

true

Exposure time: 30min

This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic

• WB

CiteAb

Western blot - Anti-RAB27A + RAB27B antibody [1G7] (AB55667)

Western Blotting using Anti-RAB27A antibody [1G7], ab55667. Publication image from Zhao, Y. et al., 2020, Nat Commun, 32024826. Legend direct from paper.

Ggpps regulates hepatocyte-derived EV secretion.a EV production by primary hepatocytes from WT and liver-specific Ggpps knockout mice (LKO) fed a normal chow diet or a HFD. (n = 6 mice per group). b CD63, TSG101 and Calnexin protein levels in parental cells and EVs from WT and LKO mice fed a normal chow diet or a HFD. c Membrane-associated Rab27A (lipid-soluble protein with geranylgeranylation in the detergent phase, down) and cytoplasm-associated Rab27A (water-soluble protein with no geranylgeranylation in the aqueous phase, up) were obtained by Triton X-114 extraction and analysed by immunoblotting. (n = 3 biologically independent samples). d Membrane-associated Rab5 (lipid-soluble protein with geranylgeranylation in the detergent phase, down) and cytoplasm-associated Rab5 (water-soluble protein with no geranylgeranylation in the aqueous phase, up) were obtained by Triton X-114 extraction and analysed by immunoblotting. (n = 3 biologically independent samples). e Fluorescence microscopy of primary hepatocytes co-infected with empty vector or Ggpps vector and WT or mutant Rab27A and stained with anti-CD63 antibodies and DAPI (nucleus, blue). (Scale bar, 5 µm). f EV production by primary hepatocytes from e. (n = 6 biologically independent samples). g TSG101 protein levels in the parental cells and EVs from e. Data from 8-week-old WT and LKO mice fed a normal chow diet or a HFD for 12 weeks are presented as the means ± SEM. Groups were analysed using an unpaired t test, *P < 0.05, **P < 0.01, ***P < 0.001, WT vs. LKO; ###P < 0.001, Chow vs. HFD. Source data are provided as a Source Data file. See also Supplementary Fig. 5.

false