ER00702

LDHA Rabbit Polyclonal Antibody

cat.: ER00702

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, FC, IF-Cell
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, 40% 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 LDHA aa1-50 / 332.
Positive control:	A431 cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, C6 cell lysate, PC-12 cell lysate, human liver tissue, mouse liver tissue, rat liver tissue, A431, RAW264.7, C6.
Subcellular location:	Cytoplasm.
Recommended Dilutions: WB IHC-P FC IF-Cell	1:5,000 1:1,000 1:1,000 1:100
Uniprot #:	SwissProt: P00338 Human \| P06151 Mouse \| P04642 Rat
Alternative names:	Cell proliferation-inducing gene 19 protein GSD11 L lactate dehydrogenase A chain L-lactate dehydrogenase A chain l7R2 Lactate dehydrogenase 1, A chain Lactate dehydrogenase A Lactate dehydrogenase A4 Lactate dehydrogenase M LDH A LDH M LDH muscle subunit LDH muscle subunit; M LDH LDH-A LDH-M LDH1 ldha LDHA_HUMAN LDHM OTTMUSP00000017774 PIG19 Proliferation-inducing gene 19 Renal carcinoma antigen NY-REN-59

Images

Fig1: Western blot analysis of LDHA on different lysates with Rabbit anti-LDHA antibody (ER00702) at 1/5,000 dilution.

Lane 1: A431 cell lysate
Lane 2: MCF7 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: RAW264.7 cell lysate
Lane 5: C6 cell lysate
Lane 6: PC-12 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 37 kDa
Observed band size: 35 kDa

Exposure time: 20 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 (ER00702) 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.

Fig2: All lanes: Western blot analysis of LDHA with anti-LDHA antibody (ER00702) at 1:500 dilution.
Lane 1: Wild-type Hela whole cell lysate (10 µg).
Lane 2/3: LDHA knockdown Hela whole cell lysate (10 µg).

ER00702 was shown to specifically react with LDHA in wild-type Hela cells. Weakened bands were observed when LDHA knockdown samples were tested. Wild-type and LDHA knockdown 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 (ER00702, 1:500) was used in 5% BSA 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-LDHA antibody (ER00702) 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 (ER00702) 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 mouse liver tissue with Rabbit anti-LDHA antibody (ER00702) 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 (ER00702) 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 rat liver tissue with Rabbit anti-LDHA antibody (ER00702) 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 (ER00702) 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: Immunocytochemistry analysis of A431 cells labeling LDHA with Rabbit anti-LDHA antibody (ER00702) 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-LDHA antibody (ER00702) 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.

Fig7: Immunocytochemistry analysis of RAW264.7 cells labeling LDHA with Rabbit anti-LDHA antibody (ER00702) 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-LDHA antibody (ER00702) 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.

Fig8: Immunocytochemistry analysis of C6 cells labeling LDHA with Rabbit anti-LDHA antibody (ER00702) 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-LDHA antibody (ER00702) 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.

Fig9: Flow cytometric analysis of A431 cells labeling LDHA.

Cells were fixed and permeabilized. Then stained with the primary antibody (ER00702, 1/1,000) (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”.