R1210-1

GAPDH Rabbit Polyclonal Antibody

cat.: R1210-1

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

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. 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: 36 kDa
Isotype:	IgG
Immunogen:	This antibody is produced by immunizing rabbits with full length recombinant protein of GAPDH.
Positive control:	HeLa cell lysate, A431 cell lysate, HEK-293 cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, C6 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, A549, LOVO, MCF-7, rat kidney tissue, human colon cancer tissue, human spleen tissue, mouse testis tissue, Hela.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:5,000-1:20,000 1:100-1:200 1:50-1:200 1:50-1:100
Uniprot #:	SwissProt: P04406 Human \| P16858 Mouse \| P46406 Rabbit
Alternative names:	38 kDa BFA-dependent ADP-ribosylation substrate aging associated gene 9 protein Aging-associated gene 9 protein BARS-38 cb609 EC 1.2.1.12 Epididymis secretory sperm binding protein Li 162eP G3P_HUMAN G3PD G3PDH GAPD GAPDH Glyceraldehyde 3 phosphate dehydrogenase Glyceraldehyde-3-phosphate dehydrogenase HEL-S-162eP KNC-NDS6 MGC102544 MGC102546 MGC103190 MGC103191 MGC105239 MGC127711 MGC88685 OCAS, p38 component OCT1 coactivator in S phase, 38-KD component peptidyl cysteine S nitrosylase GAPDH Peptidyl-cysteine S-nitrosylase GAPDH wu:fb33a10

Images

Fig1: Western blot analysis of GAPDH on different lysates with Rabbit anti-GAPDH antibody (R1210-1) at 1/20,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A431 cell lysate
Lane 3: HEK-293 cell lysate
Lane 4: Jurkat cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: RAW264.7 cell lysate
Lane 7: PC-12 cell lysate
Lane 8: C6 cell lysate
Lane 9: Mouse brain tissue lysate
Lane 10: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 36 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 (R1210-1) at 1/20,000 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: Western blot analysis of GAPDH on Oryza sativa lysates with Rabbit anti-GAPDH antibody (R1210-1) at 1/5,000 dilution.

Lysates/proteins at 15 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: 42 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 (R1210-1) 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.

Fig3: Western blot analysis of GAPDH on COS-1 cell/tissue lysates with Rabbit anti-GAPDH antibody (R1210-1) at 1/20,000 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: 2 minutes;

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 (R1210-1) at 1/20,000 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.

Fig4: Immunocytochemistry analysis of HeLa cells labeling GAPDH with Rabbit anti-GAPDH antibody (R1210-1) 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-GAPDH antibody (R1210-1) 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.

	Fig5: Immunocytochemistry analysis of NIH/3T3 cells labeling GAPDH with Rabbit anti-GAPDH antibody (R1210-1) 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-GAPDH antibody (R1210-1) 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.
	Fig6: Immunocytochemistry analysis of C6 cells labeling GAPDH with Rabbit anti-GAPDH antibody (R1210-1) 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-GAPDH antibody (R1210-1) 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: Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the antibody (R1210-1) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

	Fig8: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the antibody (R1210-1) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
	Fig9: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the antibody (R1210-1) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
	Fig10: Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the antibody (R1210-1) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
	Fig11: Flow cytometric analysis of HeLa cells labeling GAPDH. Cells were fixed and permeabilized. Then stained with the primary antibody (R1210-1, 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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