M1409-2

HADHSC Mouse Monoclonal Antibody [D10-E7]

cat.: M1409-2

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat, Zebrafish
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	D10-E7

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 34 kDa
Isotype:	IgG1
Immunogen:	Synthetic peptide within Human HADHSC aa 265-314 / 314.
Positive control:	HepG2 cell lysate, HeLa cell lysate, HT-29 cell lysate, HL-60 cell lysate, A431 cell lysate, K-562 cell lysate, human liver tissue lysate, mouse liver tissue lysate, mouse heart tissue lysate, rat liver tissue lysate, rat heart tissue lysate, zebrafish tissue lysate, HeLa, human kidney tissue, human colon carcinoma tissue, human liver tissue, mouse liver tissue, rat liver tissue.
Subcellular location:	Mitochondrion matrix.
Recommended Dilutions: WB IF-Cell IHC-P	1:1,000 1:100 1:1:1,000
Uniprot #:	SwissProt: Q16836 Human \| Q61425 Mouse \| Q9WVK7 Rat \| Q6DI22 Zebrafish
Alternative names:	3 hydroxyacyl Coenzyme A dehydrogenase HAD HADH HADH1 HADHSC HADHSC, formerly HADSC, formerly HCDH HCDH_HUMAN HHF4 Hydroxyacyl CoA dehydrogenase Hydroxyacyl-coenzyme A dehydrogenase hydroxyacyl-coenzyme A dehydrogenase, mitochondrial L 3 hydroxyacyl Coenzyme A dehydrogenase short chain M SCHAD Medium and short chain L 3 hydroxyacyl coenzyme A dehydrogenase Medium and short-chain L-3-hydroxyacyl-coenzyme A dehydrogenase MGC8392 mitochondrial MSCHAD OTTHUMP00000162626 OTTHUMP00000219688 SCHAD SCHAD, formerly Short chain 3 hydroxyacyl CoA dehydrogenase mitochondrial short chain 3-hydroxyacyl-coa dehydrogenase Short-chain 3-hydroxyacyl-CoA dehydrogenase

Images

Fig1: Western blot analysis of HADHSC on different lysates with Mouse anti-HADHSC antibody (M1409-2) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: HT-29 cell lysate (20 µg/Lane)
Lane 4: HL-60 cell lysate (20 µg/Lane)
Lane 5: A431 cell lysate (20 µg/Lane)
Lane 6: K-562 cell lysate (20 µg/Lane)
Lane 7: Human liver tissue lysate (40 µg/Lane)
Lane 8: Mouse liver tissue lysate (40 µg/Lane)
Lane 9: Mouse heart tissue lysate (40 µg/Lane)
Lane 10: Rat liver tissue lysate (40 µg/Lane)
Lane 11: Rat heart tissue lysate (40 µg/Lane)
Lane 12: Zebrafish tissue lysate (40 µg/Lane)

Predicted band size: 34 kDa
Observed band size: 34 kDa

Exposure time: 5 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 (M1409-2) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of HeLa cells labeling HADHSC with Mouse anti-HADHSC antibody (M1409-2) 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 Mouse anti-HADHSC antibody (M1409-2) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-HADHSC antibody (M1409-2) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1409-2) at 1/1,000 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Mouse anti-HADHSC antibody (M1409-2) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1409-2) at 1/1,000 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-HADHSC antibody (M1409-2) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1409-2) at 1/1,000 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.

Fig6: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Mouse anti-HADHSC antibody (M1409-2) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1409-2) at 1/1,000 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 liver tissue with Mouse anti-HADHSC antibody (M1409-2) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1409-2) at 1/1,000 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.

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