Anti-Rab 6A + Rab 6D + Ral-A 抗体 [EPR24472-24]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized A549 cells labelling Rab 6A + Rab 6D + Ral-A with ab271094 at 1/50 (11.84 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing golgi staining in A549 cell line. ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain the GM-130 at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human stomach tissue staining FUT8 with ab322558 at a 1 : 500 (1.026 ug/ml) dilution, ab271094 anti-Rab 6A + Rab 6D + Ral-A used at 1 : 2000 (0.296 ug/ml) dilution.

Panel A : merged staining of anti-FUT8 (green; Opal™520) and anti-Rab 6A + Rab 6D + Ral-A (magenta; Opal™570) on human stomach.
Panel B : anti-GUT8 showed granular staining in human stomach.
Panel C : ant-Rab 6A + Rab 6D + Ral-A showed granular staining in human stomach.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322558 and ab271094 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (Human lung carcinoma epithelial cell) cells labelling Rab 6A + Rab 6D + Ral-A with ab271094 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labelling Rab 6A + Rab 6D + Ral-A with ab271094 at 1/2000 (0.296 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Granular cytoplasmic staining on human spleen. The section was incubated with ab271094 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Rab 6A was immunoprecipitated from 0.35 mg A549 (human lung carcinoma epithelial cell) whole cell lysate with ab271094 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271094 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : A549 (human lung carcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab271094 IP in A549 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab271094 in A549 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (ab271094)

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (Mouse neuroblastoma neuroblast) cells labelling Rab 6A + Rab 6D + Ral-A with ab271094 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse colon tissue staining FUT8 with ab322558 at a 1 : 500 (1.026 ug/ml) dilution, ab271094 anti-Rab 6A + Rab 6D + Ral-A used at 1 : 2000 (0.296 ug/ml) dilution.

Panel A : merged staining of anti-FUT8 (green; Opal™520) and anti-Rab 6A + Rab 6D + Ral-A (magenta; Opal™570) on mouse colon.
Panel B : anti-GUT8 showed granular staining in mouse colon.
Panel C : ant-Rab 6A + Rab 6D + Ral-A showed granular staining in mouse colon.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322558 and ab271094 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling Rab 6A + Rab 6D + Ral-A with ab271094 at 1/2000 (0.296 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Granular cytoplasmic staining on mouse testis. The section was incubated with ab271094 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat colon tissue staining FUT8 with ab322558 at a 1 : 500 (1.026 ug/ml) dilution, ab271094 anti-Rab 6A + Rab 6D + Ral-A used at 1 : 2000 (0.296 ug/ml) dilution.

Panel A : merged staining of anti-FUT8 (green; Opal™520) and anti-Rab 6A + Rab 6D + Ral-A (magenta; Opal™570) on rat colon.
Panel B : anti-GUT8 showed granular staining in rat colon.
Panel C : ant-Rab 6A + Rab 6D + Ral-A showed granular staining in rat colon.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322558 and ab271094 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized Neuro-2a cells labelling Rab 6A + Rab 6D + Ral-A with ab271094 at 1/50 (11.84 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing golgi staining in Neuro-2a cell line. ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain GM130 at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labelling Rab 6A + Rab 6D + Ral-A with ab271094 at 1/2000 (0.296 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Granular cytoplasmic staining on rat spleen. The section was incubated with ab271094 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument .Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Rab 6A was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab271094 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271094 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate 10 ug

Lane 2 : ab271094 IP in Neuro-2a whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab271094 in Neuro-2a whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (ab271094)

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Exposure time : Lane 1-2 : 15 secondsLane 3 : 48 seconds.

All lanes:

Western blot - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (ab271094) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Lane 3:

Human kidney tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (ab271094) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 5:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 7:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 8:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 9:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Exposure time : Lane 1, 3, 4 : 48 secondsLane 2 : 3 minutes.

All lanes:

Western blot - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (ab271094) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat heart tissue lysate at 20 µg

Lane 3:

Rat kidney tissue lysate at 20 µg

Lane 4:

Rat spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Exposure time : 48 seconds.

All lanes:

Western blot - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (ab271094) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (AB271094)

Blocking and diluting buffer and concentration : 5% NFDM /TBST

All lanes:

Western blot - Anti-Rab 6A + Rab 6D + Ral-A antibody [EPR24472-24] (ab271094) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a His tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human Rab6A expression vector containing a His-tag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human Ral-A expression vector containing a His-tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 26 kDa

false

Exposure time: 1s