Anti-ALDH1A3 antibody [EPR27504-83] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ALDH1A3 antibody [EPR27504-83] - BSA and Azide free (AB308527)

This data was developed using ab308526, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HT-29 (human colorectal adenocarcinoma epithelial cell) cells labeling ALDH1A3 with ab308526 at 1/100 (5.42 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in HT-29 cell line, no staining was observed in MCF-7 cell line. Negative control : MCF-7 (PMID : 31197235). The image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-ALDH1A3 antibody [EPR27504-83] - BSA and Azide free (AB308527)

This data was developed using ab308526, the same antibody clone in a different buffer formulation.

ALDH1A3 was immunoprecipitated from 0.35 mg U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate with ab308526 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308526 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lane 2 : ab308526 IP in U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308526 in U-87 MG whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-ALDH1A3 antibody [EPR27504-83] (<a href='/en-us/products/primary-antibodies/aldh1a3-antibody-epr27504-83-ab308526'>ab308526</a>) at 1/1000 dilution

All lanes:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate

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

Observed band size: 50 kDa

false

Exposure time: 1s

• IP

Supplier Data

Immunoprecipitation - Anti-ALDH1A3 antibody [EPR27504-83] - BSA and Azide free (AB308527)

This data was developed using ab308526, the same antibody clone in a different buffer formulation.

ALDH1A3 was immunoprecipitated from 0.35 mg Mouse kidney tissue lysate with ab308526 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308526 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse kidney tissue lysate
Lane 2 : ab308526 IP in Mouse kidney tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308526 in mouse kidney tissue lysate

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

Exposure time : 128 seconds

All lanes:

Immunoprecipitation - Anti-ALDH1A3 antibody [EPR27504-83] (<a href='/en-us/products/primary-antibodies/aldh1a3-antibody-epr27504-83-ab308526'>ab308526</a>) at 1/1000 dilution

All lanes:

Mouse kidney tissue lysate

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

Observed band size: 50 kDa

false

Exposure time: 128s

• IP

Supplier Data

Immunoprecipitation - Anti-ALDH1A3 antibody [EPR27504-83] - BSA and Azide free (AB308527)

This data was developed using ab308526, the same antibody clone in a different buffer formulation.

ALDH1A3 was immunoprecipitated from 0.35 mg C6 (rat glial tumor glial cell) whole cell lysate with ab308526 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308526 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 2 : ab308526 IP in C6 (rat glial tumor glial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308526 in C6 whole cell lysate

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

Exposure time : 5 seconds

All lanes:

Immunoprecipitation - Anti-ALDH1A3 antibody [EPR27504-83] (<a href='/en-us/products/primary-antibodies/aldh1a3-antibody-epr27504-83-ab308526'>ab308526</a>) at 1/1000 dilution

All lanes:

C6 (rat glial tumor glial cell) whole cell lysate

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

Observed band size: 50 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-ALDH1A3 antibody [EPR27504-83] - BSA and Azide free (AB308527)

This data was developed using ab308526, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-ALDH1A3 antibody [EPR27504-83] (<a href='/en-us/products/primary-antibodies/aldh1a3-antibody-epr27504-83-ab308526'>ab308526</a>) at 1/1000 dilution

All lanes:

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

Observed band size: 50 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-ALDH1A3 antibody [EPR27504-83] - BSA and Azide free (AB308527)

This data was developed using ab308526, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody does not cross-react with human ALDH1A1 or ALDH1A2. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-ALDH1A3 antibody [EPR27504-83] (<a href='/en-us/products/primary-antibodies/aldh1a3-antibody-epr27504-83-ab308526'>ab308526</a>) at 1/1000 dilution

Lane 1:

His-tagged human ALDH1A3 recombinant protein at 10 ng

Lane 2:

HEK-293 transfected with a human ALDH1A1 (WT) expression vector containi a myc-His-tag®, whole cell lysate

Lane 3:

HEK-293 transfected with a human ALDH1A2 (WT) expression vector containi a myc-His-tag®, whole cell lysate

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 1/20000 dilution

Observed band size: 50 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-ALDH1A3 antibody [EPR27504-83] - BSA and Azide free (AB308527)

This data was developed using ab308526, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : MCF7 (PMID : 31197235). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : Lane 1 : 6 seconds; Lanes 2-4 : 1 second; Lane 5 : 37 seconds; Lanes 6-8 : 6 seconds.

All lanes:

Western blot - Anti-ALDH1A3 antibody [EPR27504-83] (<a href='/en-us/products/primary-antibodies/aldh1a3-antibody-epr27504-83-ab308526'>ab308526</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

Mouse kidney tissue lysate at 20 µg

Lane 6:

Rat lu tissue lysate at 20 µg

Lane 7:

Rat kidney tissue lysate at 20 µg

Lane 8:

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='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 50 kDa

false