ALDH1A3 Recombinant Rabbit Monoclonal Antibody [PSH23-87]
cat.: HA724405
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, FC, IP
Clonality: Monoclonal
Clone number: PSH23-87
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 56 kDa
Isotype: IgG
Immunogen: Recombinant protein within human ALDH1A3 aa 1-512.
Positive control: U-87 MG (Human glioblastoma cells) cell lysate, A431 (Human epidermoid carcinoma skin squamous cells) cell lysate.
Subcellular location: Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  FC
  IP
1:5,000
1:100
1:1,000
1-2μg/sample
Uniprot #: SwissProt: P47895 Human
Alternative names: Acetaldehyde dehydrogenase 6 AL1A3_HUMAN Aldehyde dehydrogenase 1A3 Aldehyde dehydrogenase 6 Aldehyde dehydrogenase family 1 member A3 Aldh1a3 ALDH1A6 ALDH6 RALDH 3 Retinaldehyde dehydrogenase 3 V1
Images
HA724405_1.jpg Fig1: Western blot analysis of ALDH1A3 on different lysates with Rabbit anti-ALDH1A3 antibody (HA724405) at 1/5,000 dilution.

Lane 1: U-87 MG (Human glioblastoma cells) cell lysate
Lane 2: A431 (Human epidermoid carcinoma skin squamous cells) cell lysate
Lane 3: MCF7 (Human breast cancer cells) cell lysate
Lane 4: 293T (Human embryonic kidney cells) cell lysate
Lane 5: Caco-2 (Human colorectal adenocarcinoma cells) cell lysate
Lane 6: HeLa (Human cervical adenocarcinoma cells) cell lysate

Lysates/proteins at 10 µg/Lane.
Exposure time: 30 seconds; ECL: K1801

Low expression: MCF7 cells (PMID: 31197235).
Low expression of ALDH1A3 protein in 293T, Caco-2, and HeLa cells is consistent with the predicted expression pattern.

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724405, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 56 kDa
Observed band size: 50-56 kDa
HA724405_2.jpg Fig2: Western blot analysis of ALDH1A3 on different lysates with Rabbit anti-ALDH1A3 antibody (HA724405) at 1/5,000 dilution.

Lane 1: Human ALDH1A3 recombinant protein
Lane 2: Human ALDH1A2 recombinant protein
Lane 3: Human ALDH1A1 recombinant protein

Lysates/proteins at 20 ng/Lane.
Exposure time: 7 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724405, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted MW: 56 kDa
Observed MW: 58-61 kDa
HA724405_3.jpg Fig3: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: U-87 MG (Human glioblastoma cells)/MCF7 (Human breast cancer cells)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA724405, 1/100, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).
Low expression: MCF7 cell (PMID: 31197235).
HA724405_4.jpg Fig4: Application: Flow Cytometry (Intra)

Species: Human
Sample: U-87 MG (Human glioblastoma cells)/MCF7 (Human breast cancer cells)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724405 (1/1,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Low expression: MCF7 cell (PMID: 31197235).
HA724405_5.jpg Fig5: Immunoprecipitation (IP)

ALDH1A3 was immunoprecipitated in 0.2 mg U-87 MG (Human glioblastoma cells) cell lysate with HA724405 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724405 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: U-87 MG cell lysate (input)
Lane 2: HA724405 IP in U-87 MG cell lysate
Lane 3: Rabbit IgG instead of HA724405 in U-87 MG cell lysate

Exposure time: 14 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724405, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 56 kDa
Observed band size: 50-56 kDa
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.