HA721733

Aldolase A/B/C Recombinant Rabbit Monoclonal Antibody [PSH01-81]

cat.: HA721733

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IHC-Fr, IF-Tissue, IF-Cell
Clonality:	Monoclonal
Clone number:	PSH01-81

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 40 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Aldolase C aa 50-100.
Positive control:	U-87 MG cell lysate, RAW264.7 cell lysate, C6 cell lysate, human brain tissue lysate, mouse brain tissue lysate, mouse pancreas tissue lysate, rat brain tissue lysate, rat pancreas tissue lysate, U-87 MG, mouse brain tissue, rat brain tissue.
Subcellular location:	cytoskeleton, cytosol, extracellular exosome, extracellular region
Recommended Dilutions: WB IHC-P IHC-Fr IF-Tissue IF-Cell	1:2,000-1:5,000 1:20,000 1:1,000 1:1,000 1:100
Uniprot #:	SwissProt: P09972 Human \| P05063 Mouse \| P09117 Rat
Alternative names:	ALDC ALDO C aldoc ALDOC_HUMAN Aldolase 3 Aldolase C Fructose bisphosphate Brain type aldolase Brain-type aldolase Fructoaldolase C Fructose 1 6 biphosphate triosephosphate lyase Fructose bisphosphate aldolase C Fructose-bisphosphate aldolase C

Images

	Fig1: Immunofluorescence analysis of frozen mouse brain tissue with Rabbit anti-Aldolase A/B/C antibody (HA721733) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721733, green) at 1/1,000 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig2: Immunofluorescence analysis of frozen rat brain tissue with Rabbit anti-Aldolase A/B/C antibody (HA721733) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721733, green) at 1/1,000 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig3: Western blot analysis of Aldolase A/B/C on different lysates with Rabbit anti-Aldolase A/B/C antibody (HA721733) at 1/5,000 dilution. Lane 1: U-87 MG-si NT cell lysate (10 µg/Lane) Lane 2: U-87 MG-si Aldolase A/B/C cell lysate (10 µg/Lane) Predicted band size: 40 kDa Observed band size: 40 kDa Exposure time: 4 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721733) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig4: Western blot analysis of Aldolase A/B/C on different lysates with Rabbit anti-Aldolase A/B/C antibody (HA721733) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: U-87 MG cell lysate
Lane 2: RAW264.7 cell lysate
Lane 3: C6 cell lysate
Lane 4: Human brain tissue lysate
Lane 5: Mouse brain tissue lysate
Lane 6: Mouse pancreas tissue lysate (negative)
Lane 7: Rat brain tissue lysate
Lane 8: Rat pancreas tissue lysate (negative)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 40 kDa
Observed band size: 40 kDa

Exposure time: 19 seconds;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721733) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig5: Immunocytochemistry analysis of U-87 MG cells labeling Aldolase A/B/C with Rabbit anti-Aldolase A/B/C antibody (HA721733) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Aldolase A/B/C antibody (HA721733) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig6: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Aldolase A/B/C antibody (HA721733) at 1/20,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721733) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig7: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Aldolase A/B/C antibody (HA721733) at 1/20,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721733) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig8: Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling Aldolase A/B/C with Rabbit anti-Aldolase A/B/C antibody (HA721733) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721733, green) at 1/1,000 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.