HA721012

RAB5C Recombinant Rabbit Monoclonal Antibody [JE63-14]

cat.: HA721012

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell
Clonality:	Monoclonal
Clone number:	JE63-14

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: 23 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human RAB5C aa 1-216/216.
Positive control:	HAP1-parental cell lysate, HAP1-商品名 KD cell lysate, RAW264.7 cell lysate, Neuro-2a cell lysate, MEF cell lysate, NR8383 cell lysate, 293 cell lysate, HepG2 cell lysate, LOVO cell lysate, SK-Br-3 cell lysate, NR8383, HepG2, human kidney tissue, mouse kidney tissue, rat kidney tissue.
Subcellular location:	Early endosome membrane, Cell membrane, Melanosome.
Recommended Dilutions: WB IHC-P IF-Cell	1:1,000-1:5,000 1:1,000 1:500
Uniprot #:	SwissProt: P51148 Human \| P35278 Mouse Entrez Gene: 287709 Rat
Alternative names:	L1880 RAB, member of RAS oncogene family like RAB5C RAB5C, member of RAS oncogene family RAB5C, member RAS oncogene family RAB5C_HUMAN RAB5CL RAB5L RABL Ras-related protein Rab-5C

Images

Fig1: Western blot analysis of RAB5C on different lysates with Rabbit anti-RAB5C antibody (HA721012) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-RAB5C KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 23 kDa
Observed band size: 23 kDa

Exposure time: 30 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 (HA721012) at 1/1,000 dilution was used in K1803 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 RAB5C on different lysates with Rabbit anti-RAB5C antibody (HA721012) at 1/5,000 dilution.

Lane 1: RAW264.7 cell lysate
Lane 2: Neuro-2a cell lysate
Lane 3: MEF cell lysate
Lane 4: NR8383 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 23 kDa
Observed band size: 23 kDa

Exposure time: 6 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 (HA721012) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 RAB5C on different lysates with Rabbit anti-RAB5C antibody (HA721012) at 1/1,000 dilution.

Lane 1: 293 cell lysate
Lane 2: HepG2 cell lysate
Lane 3: LOVO cell lysate
Lane 4: SK-Br-3 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 23 kDa
Observed band size: 23 kDa

Exposure time: 1 minute;

15% 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 (HA721012) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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 NR8383 cells labeling RAB5C with Rabbit anti-RAB5C antibody (HA721012) at 1/500 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-RAB5C antibody (HA721012) 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 (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.

	Fig5: Immunocytochemistry analysis of HepG2 cells labeling RAB5C with Rabbit anti-RAB5C antibody (HA721012) at 1/500 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-RAB5C antibody (HA721012) 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 (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.
	Fig6: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-RAB5C antibody (HA721012) 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 (HA721012) 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-RAB5C antibody (HA721012) 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 (HA721012) 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 rat kidney tissue with Rabbit anti-RAB5C antibody (HA721012) 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 (HA721012) 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.

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