0807-1

Lactate Dehydrogenase B Rabbit Polyclonal Antibody

cat.: 0807-1

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, FC, IHC-P
Clonality:	Polyclonal

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, 25% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 37 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human LDH-B2 aa 285-334 / 334.
Positive control:	Hela cell lysate, Jurkat cell lysate, Daudi cell lysate, HL-60 cell lysate, A549, rat kidney tissue, human kidney tissue, mouse kidney tissue.
Subcellular location:	Cytoplasm, Membrane, Mitochondrion, Mitochondrion inner membrane.
Recommended Dilutions: WB IF-Cell FC IHC-P	1:500-1:1,000 1:200 1:500-1:1,000 1:200-1:500
Uniprot #:	SwissProt: P07195 Human \| P16125 Mouse \| P42123 Rat
Alternative names:	Epididymis secretory protein Li 281 HEL S 281 L lactate dehydrogenase B chain L-lactate dehydrogenase B chain Lactate dehydrogenase H chain LDH B LDH H LDH heart subunit LDH-B LDH-H LDHB LDHB_HUMAN LDHBD LDHH Renal carcinoma antigen NY REN 46 Renal carcinoma antigen NY-REN-46 TRG-5 TRG5

Images

Fig1: Western blot analysis of Lactate Dehydrogenase B on different lysates with Rabbit anti-Lactate Dehydrogenase B antibody (0807-1) at 1/500 dilution.

Lane 1: Hela cell lysate
Lane 2: Jurkat cell lysate
Lane 3: Daudi cell lysate
Lane 4: HL-60 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 37 kDa
Observed band size: 37 kDa

Exposure time: 2 minutes;

12% 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 (0807-1) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of Lactate Dehydrogenase on different lysates with Rabbit anti-Lactate Dehydrogenase antibody (0807-1) at 1/1,000 dilution.

Lane 1: Hela-si NT cell lysate
Lane 2: Hela-si Lactate Dehydrogenase cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 37 kDa
Observed band size: 37 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

0807-1 was shown to specifically react with Lactate Dehydrogenase in Hela-si NT cells. Weakened band was observed when Hela-si Lactate Dehydrogenase sample was tested. Hela-si NT and Hela-si Lactate Dehydrogenase samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (0807-1, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of A549 cells labeling Lactate Dehydrogenase B with Rabbit anti-Lactate Dehydrogenase B antibody (0807-1) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Lactate Dehydrogenase B antibody (0807-1) at 1/200 dilution in 2% negative goat serum 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Lactate Dehydrogenase B antibody (0807-1) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (0807-1) at 1/200 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-Lactate Dehydrogenase B antibody (0807-1) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (0807-1) at 1/200 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 kidney tissue with Rabbit anti-Lactate Dehydrogenase B antibody (0807-1) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (0807-1) at 1/200 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: Flow cytometric analysis of A549 cells labeling Lactate Dehydrogenase B.

Cells were fixed and permeabilized. Then stained with the primary antibody (0807-1, 1ug/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).

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