HA722901

SDHB Recombinant Rabbit Monoclonal Antibody [JE77-70]

cat.: HA722901

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IP, IF-Cell
Clonality:	Monoclonal
Clone number:	JE77-70

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. 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: 32 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide.
Positive control:	K-562 cell lysate, HEK-293 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse liver tissue lysate, Rat liver tissue lysate, K-562, NIH/3T3, human liver cancer tissue, human kidney tissue, rat kidney tissue, mouse kidney tissue, PC-12.
Subcellular location:	Mitochondrion inner membrane.
Recommended Dilutions: WB IHC-P IP IF-Cell	1:2,000 1:1,000 1-2μg/sample 1:250
Uniprot #:	SwissProt: P21912 Human \| Q9CQA3 Mouse \| P21913 Rat
Alternative names:	CWS2 DHSB_HUMAN FLJ92337 Ip Iron sulfur subunit Iron sulfur subunit of complex II Iron-sulfur subunit of complex II mitochondrial PGL 4 PGL4 SDH 1 SDH SDH1 SDH2 SDH2, homolog of SdhB SDHIP Succinate dehydrogenase [ubiquinone] iron sulfur protein mitochondrial Succinate dehydrogenase [ubiquinone] iron sulfur subunit Succinate dehydrogenase [ubiquinone] iron-sulfur subunit succinate dehydrogenase [ubiquinone] iron-sulfur subunit, mitochondrial Succinate Dehydrogenase 1 Iron Sulfur Subunit Succinate Dehydrogenase 2, S. cerevisiae, homolog of Succinate dehydrogenase complex iron sulfur subunit B Succinate dehydrogenase complex subunit B iron sulfur Succinate Dehydrogenase Complex Subunit B Iron Sulfur Protein succinate dehydrogenase complex, subunit B, iron sulfur (Ip) Succinate dehydrogenase iron sulfur protein

Images

Fig1: Western blot analysis of SDHB on different lysates with Rabbit anti-SDHB antibody (HA722901) at 1/2,000 dilution.

Lane 1: K-562 cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: PC-12 cell lysate
Lane 5: Mouse liver tissue lysate
Lane 6: Rat liver tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 32 kDa
Observed band size: 30 kDa

Exposure time: 4 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 (HA722901) at 1/2,000 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 K-562 cells labeling SDHB with Rabbit anti-SDHB antibody (HA722901) at 1/250 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-SDHB antibody (HA722901) at 1/250 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 (HA601187, 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 NIH/3T3 cells labeling SDHB with Rabbit anti-SDHB antibody (HA722901) at 1/250 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-SDHB antibody (HA722901) at 1/250 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 (HA601187, 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-SDHB antibody (HA722901) 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 (HA722901) 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 kidney tissue with Rabbit anti-SDHB antibody (HA722901) 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 (HA722901) 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 rat kidney tissue with Rabbit anti-SDHB antibody (HA722901) 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 (HA722901) 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 mouse kidney tissue with Rabbit anti-SDHB antibody (HA722901) 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 (HA722901) 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: SDHB was immunoprecipitated from 0.2 mg HEK-293 cell lysate with HA722901 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722901 at 1/1,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody (M1003-7) at 1/100,000 dilution was used for 1 hour at room temperature. Lane 1: HEK-293 cell lysate (input) Lane 2: HA722901 IP in HEK-293 cell lysate Lane 3: Rabbit IgG instead of HA722901 in HEK-293 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 40 seconds; ECL: K1801

Fig9: Immunocytochemistry analysis of PC-12 cells labeling SDHB with Rabbit anti-SDHB antibody (HA722901) at 1/250 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-SDHB antibody (HA722901) at 1/250 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.

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