Anti-RAB35 antibody [EPR24487-21] (BSA and Azide free)

• IP

Lab

Immunoprecipitation - Anti-RAB35 antibody [EPR24487-21] (BSA and Azide free) (AB300117)

This data was developed using ab300116, the same antibody clone in a different buffer formulation.

RAB35 was immunoprecipitated from 0.35 mg Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate 10 µg with ab300116 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300116 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

All lanes:

Immunoprecipitation - Anti-RAB35 antibody [EPR24487-21] (<a href='/en-us/products/primary-antibodies/rab35-antibody-epr24487-21-ab300116'>ab300116</a>) at 1/1000 dilution

Lane 1:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/rab35-antibody-epr24487-21-ab300116'>ab300116</a> at 1/30 IP in Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/rab35-antibody-epr24487-21-ab300116'>ab300116</a> in Caco-2 whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 32s

• IP

Supplier Data

Immunoprecipitation - Anti-RAB35 antibody [EPR24487-21] (BSA and Azide free) (AB300117)

This data was developed using ab300116, the same antibody clone in a different buffer formulation.

Immunoprecipitation of RAB35 in HAP1 cells. Lysates were prepared and immunoprecipitation was performed using 2 μg of ab300116 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

This data was 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:

Immunoprecipitation - Anti-RAB35 antibody [EPR24487-21] (<a href='/en-us/products/primary-antibodies/rab35-antibody-epr24487-21-ab300116'>ab300116</a>) at 2 µg

false

• WB

Supplier Data

Western blot - Anti-RAB35 antibody [EPR24487-21] (BSA and Azide free) (AB300117)

This data was developed using ab300116, the same antibody clone in a different buffer formulation.

ab300116 was shown to react with RAB35 in wild-type HAP1 cells in Western blot with loss of signal observed in a RAB35 knockout cell line. Wild-type HAP1 and RAB35 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab300116 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was 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.

Lanes 1 - 2:

Western blot - Anti-RAB35 antibody [EPR24487-21] (<a href='/en-us/products/primary-antibodies/rab35-antibody-epr24487-21-ab300116'>ab300116</a>) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Anti-RAB35 antibody [EPR24487-21] (BSA and Azide free) (ab300117) at 1/1000 dilution

Lane 1:

Wild-type HAP1 lysate at 20 µg

Lane 2:

RAB35 knock-out HAP1 lysate at 20 µg

false

• WB

Supplier Data

Western blot - Anti-RAB35 antibody [EPR24487-21] (BSA and Azide free) (AB300117)

This data was developed using ab300116, the same antibody clone in a different buffer formulation.

This antibody has no cross-reactivity with other RAB family proteins.

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

All lanes:

Western blot - Anti-RAB35 antibody [EPR24487-21] (<a href='/en-us/products/primary-antibodies/rab35-antibody-epr24487-21-ab300116'>ab300116</a>) at 1/1000 dilution

Lane 1:

RAB35 full-length protein at 10 ng

Lane 2:

RAB family protein homologous region at 10 ng

Secondary

All lanes:

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 1/100000 dilution

Predicted band size: 23 kDa

Observed band size: 25 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-RAB35 antibody [EPR24487-21] (BSA and Azide free) (AB300117)

This data was developed using ab300116, the same antibody clone in a different buffer formulation.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29662076; PMID : 34349123).

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

All lanes:

Western blot - Anti-RAB35 antibody [EPR24487-21] (<a href='/en-us/products/primary-antibodies/rab35-antibody-epr24487-21-ab300116'>ab300116</a>) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Secondary

All lanes:

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

Predicted band size: 23 kDa

Observed band size: 25 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-RAB35 antibody [EPR24487-21] (BSA and Azide free) (AB300117)

This data was developed using ab300116, the same antibody clone in a different buffer formulation.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 23572513).

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

All lanes:

Western blot - Anti-RAB35 antibody [EPR24487-21] (<a href='/en-us/products/primary-antibodies/rab35-antibody-epr24487-21-ab300116'>ab300116</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

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

Secondary

All lanes:

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

Predicted band size: 23 kDa

Observed band size: 25 kDa

false

Exposure time: 3min