Anti-Rab3A antibody [EPR24450-44] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-Rab3A antibody [EPR24450-44] - BSA and Azide free (AB285170)

This data was developed using ab285163, the same antibody clone in a different buffer formulation.

Rab3A was immunoprecipitated from 0.35 mg Human eyeball tissue lysate 10 ug with ab285163 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab285163 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Human eyeball tissue lysate 10 ug

Lane 2 : ab285163 IP in Human eyeball tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab285163 in human eyeball tissue lysate

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

Exposure time : 27 seconds

All lanes:

Immunoprecipitation - Anti-Rab3A antibody [EPR24450-44] (<a href='/en-us/products/primary-antibodies/rab3a-antibody-epr24450-44-ab285163'>ab285163</a>)

Predicted band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Rab3A antibody [EPR24450-44] - BSA and Azide free (AB285170)

This data was developed using ab285163, the same antibody clone in a different buffer formulation.

Lane 1 : His-tagged human RAB3A recombinant protein(aa1-220)
Lane 2 : His-tagged human RAB3B recombinant protein(aa1-219)
Lane 3 : His-tagged human RAB3C recombinant protein(aa1-227)
Lane 4 : His-tagged human RAB3D recombinant protein(aa1-219)

A band around 55 kDa may be RAB3A dimer.

All lanes:

Western blot - Anti-Rab3A antibody [EPR24450-44] (<a href='/en-us/products/primary-antibodies/rab3a-antibody-epr24450-44-ab285163'>ab285163</a>) at 1/1000 dilution

Lane 1:

His-tagged human RAB3A recombinant protein

Lane 2:

His-tagged human RAB3B recombinant protein

Lane 3:

His-tagged human RAB3C recombinant protein

Lane 4:

His-tagged human RAB3D recombinant protein

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Rab3A antibody [EPR24450-44] - BSA and Azide free (AB285170)

This data was developed using ab285163, the same antibody clone in a different buffer formulation.

Negative control : Human skeletal muscle (PMID : 11865063).

Exposure time : 5.5 seconds

All lanes:

Western blot - Anti-Rab3A antibody [EPR24450-44] (<a href='/en-us/products/primary-antibodies/rab3a-antibody-epr24450-44-ab285163'>ab285163</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate

Lane 2:

Human skeletal muscle tissue lysate

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Rab3A antibody [EPR24450-44] - BSA and Azide free (AB285170)

This data was developed using ab285163, the same antibody clone in a different buffer formulation.

ab285163 was shown to react with RAB3A in wild-type SKNFI cells in Western blot with loss of signal observed in a RAB3A knockout cell line. Wild-type SKNFI and RAB3A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab285163 overnight at 4 °C at a 1/200 dilution. Blots were incubated with secondary antibodies at 0.2 µg/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-Rab3A antibody [EPR24450-44] (<a href='/en-us/products/primary-antibodies/rab3a-antibody-epr24450-44-ab285163'>ab285163</a>) at 1/200 dilution

Lane 1:

Wild-type SKNFI lysate at 40 µg

Lane 2:

RAB3A knock-out SKNFI lysate at 40 µg

false