Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300509, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling ALDH1L1+ALDH1L2 with ab300509 at 1/50 (10.72 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 HepG2 cell line is observed. 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-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300510, the same antibody clone in a different buffer formulation. ALDH1L1+ALDH1L2 was immunoprecipitated from 0.35 mg Human liver tissue lysate 10 ug with ab300509 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300509 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Human liver tissue lysate 10 ug Lane 2 : ab300509 IP in Human liver tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300509 in human liver tissue lysate. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 3 seconds

Lane 1:

Immunoprecipitation - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/aldh1l1aldh1l2-antibody-epr25443-54-astrocyte-marker-ab300509'>ab300509</a>) at 1/30 dilution

Lane 2:

Immunoprecipitation - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/aldh1l1aldh1l2-antibody-epr25443-54-astrocyte-marker-ab300509'>ab300509</a>) at 1/1000 dilution

All lanes:

Human liver tissue 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: 3s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300509, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling ALDH1L1+ALDH1L2 with ab300509 at 1/5000 (0.107 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on astrocytes of mouse cerebrum. The section was incubated with ab300509 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) was used.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-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300509, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling ALDH1L1+ALDH1L2 with ab300509 at 1/500 (1.072 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on astrocytes of rat cerebrum (PMID : 18171944). The section was incubated with ab300509 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) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300509, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron/glia cell cells labelling ALDH1L1+ALDH1L2 with ab300509 at 1/50 (10.72 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 mouse primary astrocyte.Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. is observed. Anti-Glial Fibrillary Acidic Protein (GFAP) mouse monoclonal antibody was used to counterstain tubulin at 1/50 10ug/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.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300509, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum tissue labeling ALDH1L1+ALDH1L2 with ab300509 at 1/100 (5.36 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Positive staining on rat cerebrum is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300509, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neuron/glia cell cells labelling ALDH1L1+ALDH1L2 with ab300509 at 1/50 (10.72 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2µg/mL dilution (Green). Confocal image showing cytoplasmic staining in rat primary astrocyte. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. is observed. Anti-Glial Fibrillary Acidic Protein (GFAP) mouse monoclonal antibody was used to counterstain tubulin at 1/50 10µg/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 2µg/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300509, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling ALDH1L1+ALDH1L2 with ab300509 at 1/5000 (0.107 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse liver. The section was incubated with ab300509 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) was used.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-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300509, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cell cells labelling ALDH1L1+ALDH1L2 with ab300509 at 1/50 dilution (1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300509, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum tissue labeling ALDH1L1+ALDH1L2 with ab300509 at 1/100 (5.36 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IP

Supplier Data

Immunoprecipitation - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300510, the same antibody clone in a different buffer formulation. ALDH1L1+ALDH1L2 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate 10 ug with ab300509 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300509 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse brain tissue lysate 10 ug Lane 2 : ab300509 IP in Mouse brain tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300509 in mouse brain tissue lysate. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 32 seconds

Lane 1:

Immunoprecipitation - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/aldh1l1aldh1l2-antibody-epr25443-54-astrocyte-marker-ab300509'>ab300509</a>) at 1/30 dilution

Lane 2:

Immunoprecipitation - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/aldh1l1aldh1l2-antibody-epr25443-54-astrocyte-marker-ab300509'>ab300509</a>) at 1/1000 dilution

All lanes:

Mouse brain tissue 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

• WB

Supplier Data

Western blot - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300509, the same antibody clone in a different buffer formulation. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : Lane 1 : 6 seconds; lane 2 : 37 seconds; lane 3 : 15 seconds; Lane 4 : 37 seconds; lanes 5-6 : 8 seconds; lane 7 : 59 seconds. Bands below 100 kDa may due to degradation (PMID : 29979702).

All lanes:

Western blot - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/aldh1l1aldh1l2-antibody-epr25443-54-astrocyte-marker-ab300509'>ab300509</a>) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Lane 5:

Rat liver tissue lysate at 20 µg

Lane 6:

C8-D1A (mouse cerebellum astrocyte) whole cell lysate at 20 µg

Lane 7:

PC-12 (rat adrenal gland pheochromocytoma) 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 1/100000 dilution

Observed band size: 100 kDa

false

• WB

Supplier Data

Western blot - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 3 seconds.

All lanes:

Western blot - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/aldh1l1aldh1l2-antibody-epr25443-54-astrocyte-marker-ab300509'>ab300509</a>) at 1/5000 dilution

Observed band size: 100 kDa

false

Exposure time: 3s

• Dot

Supplier Data

Dot Blot - Anti-ALDH1L1+ALDH1L2 antibody [EPR25443-54] - BSA and Azide free (AB300510)

This data was developed using ab300509, the same antibody clone in a different buffer formulation.  Dot blot analysis of ALDH1L1 and ALDH1L2 using ab300509 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.  Lane1 : His-tagged recombinant mouse ALDH1L1 protein Lane 2 : His-tagged recombinant mouse ALDH1L2 protein  Blocking and diluting buffer and concentration : 5% NFDM/TBST.  Exposure time : 48 seconds.