Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) - BSA and Azide free (AB307608)

This data was developed using ab307607, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol-permeabilized RAB8A KO HeLa (ED010027, Left) / Parental HeLa (EDWT01, Right) cells labeling RAB8A with ab307607 at 1/800 dilution (0.1 ug) (Red) compared with a mouse monoclonal IgG (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.

• IHC

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Immunohistochemistry - Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) - BSA and Azide free (AB307608)

This data was developed using ab307607, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human breast tissue labeling RAB8A with ab307607 at 1/5000 dilution (0.167 µg/mL) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on human breast. The section was incubated with ab307607 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC

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Immunohistochemistry - Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) - BSA and Azide free (AB307608)

This data was developed using ab307607, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded cell pellets labeling RAB8A with ab307607 at 1/5000 dilution (0.167 µg/mL) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on (A) Wild-type HeLa cell pellets, no staining on (B) RAB8A knockout HeLa (ab264993) cell pellets. The section was incubated with ab307607 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC

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Immunohistochemistry - Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) - BSA and Azide free (AB307608)

This data was developed using ab307607, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human bladder carcinoma tissue labeling RAB8A with ab307607 at 1/5000 dilution (0.167 µg/mL) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on human bladder carcinoma. The section was incubated with ab307607 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) - BSA and Azide free (AB307608)

This data was developed using ab307607, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma epithelial cell) cells labeling RAB8A with ab307607 at 1/500 dilution, followed by ab98712 Goat Anti-Mouse IgG Fc (DyLight® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml) (Green). Confocal image showing cytoplasmic staining in A549 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab202272 Recombinant Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab202272) was used to counterstain tubulin at 1/50 dilution (10 ug/ml) (Red). Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab98712 Goat Anti-Mouse IgG Fc (DyLight® 488) preadsorbed at 1/1000 dilution (2 µg/ml).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) - BSA and Azide free (AB307608)

This data was developed using ab307607, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAB8A KO HeLa (RAB8A knockout human cervical adenocarcinoma epithelial cell) (ab264993) cells labeling RAB8A with ab307607 at 1/500 dilution, followed by ab98712 Goat Anti-Mouse IgG Fc (DyLight® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml) (Green). Confocal image showing cytoplasmic staining in Parental HeLa cell line, and no staining in RAB8A KO HeLa cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab202272 Recombinant Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab202272) was used to counterstain tubulin at 1/50 dilution (10 µg/ml) (Red). Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab98712 Goat Anti-Mouse IgG Fc (DyLight® 488) preadsorbed at 1/1000 dilution (2 µg/ml).

• IP

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Immunoprecipitation - Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) - BSA and Azide free (AB307608)

This data was developed using ab307607, the same antibody clone in a different buffer formulation. RAB8A was immunoprecipitated from 0.35 mg A549 (human lung carcinoma epithelial cell) whole cell lysate 10 ug with ab307607 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307607 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : A549 whole cell lysate 10 ug Lane 2 : ab307607 IP in A549 whole cell lysate Lane 3 : Mouse IgG2a monoclonal isotype control (ab18413) instead of ab307607 in A549 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 24 seconds.

All lanes:

Immunoprecipitation - Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/rab8a-antibody-mjf-r22-79-3-mouse-igg2a-chimeric-ab307607'>ab307607</a>) at 1/1000 dilution

Lane 1:

A549 (human lung carcinoma epithelial cell) whole cell lysate 10 μg

Lane 2:

A549 whole cell lysate

Observed band size: 23 kDa

false

Exposure time: 24s

• WB

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Western blot - Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) - BSA and Azide free (AB307608)

This data was developed using ab307607, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, ab307607 was shown to bind specifically to RAB8A. A band was observed at 23 kDa in wild-type HeLa cell lysates with whereas no signal observed at this size in RAB8A knockout cell line ab264993 (knockout cell lysate ab257195). Exposure time : 48 seconds.

All lanes:

Western blot - Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/rab8a-antibody-mjf-r22-79-3-mouse-igg2a-chimeric-ab307607'>ab307607</a>) at 1/5000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 20 μg

Lane 2:

RAB8A knockout HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 20 μg

Lane 3:

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

Lane 4:

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

Observed band size: 23 kDa

false

Exposure time: 48s