MA5-15692

antibody from Invitrogen Antibodies

Targeting: ALDH1A1 ALDH1, PUMB1, RALDH1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15692

Provider

Invitrogen Antibodies

Product name

ALDH1A1 Monoclonal Antibody (5A11)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

MA5-15692 targets ALDH1A1 in IHC and WB applications and shows reactivity with Human samples.

Antibody clone number

5A11

Concentration

Conc. Not Determined

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of ALDH1A1 using ALDH1A1 monoclonal antibody (Product # MA5-15692) in Raji (1), Jurkat (2), THP-1 (3) and K562 (4) cell lysate.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-ALDH1A1 Monoclonal Antibody (5A11)(Product # MA5-15692) and a 55kDa band corresponding to ALDH1A1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1), Hep G2 (Lane 2), MCF7 (Lane 3), SK-O-V3 (Lane 4), NIH:OVCAR-3 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).Expression of ALDH1A1 was found to be higher in A549,Hep G2 and NIH:OVCAR-3 as compared to MCF-7 and SK-O-V3.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of ALDH1A1 was performed using 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with ALDH1A1 Monoclonal Antibody (5A11) (Product # MA5-15692) at 5 µg in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing Cytoplasm localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60x magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded cerebellum tissues (left) and lung cancer (right) using ALDH1A1 monoclonal antibody (Product # MA5-15692) followed with DAB staining.