ET1703-40

SDHA Recombinant Rabbit Monoclonal Antibody [JM10-83]

cat.: ET1703-40

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Zebrafish
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JM10-83

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 73 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human SDHA aa 581-624 / 664.
Positive control:	Jurkat cell lysate, HL-60 cell lysate, HeLa cell lysate, MCF7 cell lysate, C2C12 cell lysate, L6 cell lysate, Mouse brain tissue lysate, Mouse heart tissue lysate, Rat brain tissue lysate, Rat heart tissue lysate, Zebrafish tissue lysate, HeLa, human kidney tissue, C2C12, human heart tissue, human liver tissue, mouse colon tissue, mouse skeletal muscle tissue, mouse testis tissue, mouse kidney tissue, rat kidney tissue, rat heart tissue, zebrafish tissue.
Subcellular location:	Mitochondrion inner membrane.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:500-1:1,000 1:100-1:500 1:200-1:1,000 1:1,000-1:5,000 1:1,000
Uniprot #:	SwissProt: P31040 Human \| Q8K2B3 Mouse \| Q920L2 Rat
Alternative names:	CMD1GG DHSA_HUMAN Flavoprotein subunit of complex II Fp PGL5 SDH 2 SDH1 SDH2 SDHA SDHF Succinate dehydrogenase [ubiquinone] flavoprotein subunit Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial Succinate dehydrogenase [ubiquinone] flavoprotein subunit, mitochondrial Succinate dehydrogenase complex flavoprotein subunit A Succinate dehydrogenase complex flavoprotein subunit Succinate dehydrogenase complex flavoprotein subunit precursor Succinate dehydrogenase complex subunit A Succinate dehydrogenase complex subunit A flavoprotein (Fp) Succinate dehydrogenase complex subunit A flavoprotein

Images

Fig1: Western blot analysis of SDHA on different lysates with Rabbit anti-SDHA antibody (ET1703-40) at 1/1,000 dilution.

Lane 1: Jurkat cell lysate (15 µg/Lane)
Lane 2: HL-60 cell lysate (15 µg/Lane)
Lane 3: HeLa cell lysate (15 µg/Lane)
Lane 4: MCF7 cell lysate (15 µg/Lane)
Lane 5: C2C12 cell lysate (15 µg/Lane)
Lane 6: L6 cell lysate (15 µg/Lane)
Lane 7: Mouse brain tissue lysate (30 µg/Lane)
Lane 8: Mouse heart tissue lysate (30 µg/Lane)
Lane 9: Rat brain tissue lysate (30 µg/Lane)
Lane 10: Rat heart tissue lysate (30 µg/Lane)
Lane 11: Zebrafish tissue lysate (30 µg/Lane)

Predicted band size: 73 kDa
Observed band size: 70 kDa

Exposure time: 5 seconds; ECL: K1801;

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 (ET1703-40) at 1/1,000 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.

Fig2: Immunocytochemistry analysis of HeLa cells labeling SDHA with Rabbit anti-SDHA antibody (ET1703-40) at 1/500 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-SDHA antibody (ET1703-40) at 1/500 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.

	Fig3: Immunocytochemistry analysis of C2C12 cells labeling SDHA with Rabbit anti-SDHA antibody (ET1703-40) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-SDHA antibody (ET1703-40) 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.
	Fig4: Immunofluorescence analysis of paraffin-embedded human kidney tissue labeling SDHA with Rabbit anti-SDHA antibody (ET1703-40) at 1/200 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 (ET1703-40, green) at 1/200 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).
	Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-SDHA antibody (ET1703-40) at 1/5,000 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 (ET1703-40) at 1/5,000 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 human heart tissue with Rabbit anti-SDHA antibody (ET1703-40) at 1/2,000 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 (ET1703-40) at 1/2,000 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 human liver tissue with Rabbit anti-SDHA antibody (ET1703-40) 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 (ET1703-40) 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-SDHA antibody (ET1703-40) 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 (ET1703-40) 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.

	Fig9: Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue with Rabbit anti-SDHA antibody (ET1703-40) 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 (ET1703-40) 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.
	Fig10: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-SDHA antibody (ET1703-40) 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 (ET1703-40) 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.
	Fig11: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-SDHA antibody (ET1703-40) at 1/5,000 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 (ET1703-40) at 1/5,000 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.

	Fig12: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-SDHA antibody (ET1703-40) at 1/5,000 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 (ET1703-40) at 1/5,000 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.
	Fig13: Immunohistochemical analysis of paraffin-embedded rat heart tissue with Rabbit anti-SDHA antibody (ET1703-40) at 1/2,000 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 (ET1703-40) at 1/2,000 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.
	Fig14: Immunohistochemical analysis of paraffin-embedded zebrafish tissue with Rabbit anti-SDHA antibody (ET1703-40) 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 (ET1703-40) 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.

Fig15: Flow cytometric analysis of HeLa cells labeling SDHA.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1703-40, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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