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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [5]
- Immunocytochemistry [2]
- Immunohistochemistry [8]
- Other assay [1]
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- Product number
- PA5-27522 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- hnRNP K Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, Hela, HepG2, Mouse brain.
- Concentration
- 0.18 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of hnRNP K using 30 µg of A) 293T (B) A431 and C) H1299 lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with a hnRNP K polyclonal antibody (Product # PA5-27522) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), PANC-1 (Lane 2), MCF7 (Lane 3), U-2 OS (Lane 4), U-87 MG (Lane 5), IMR32 (Lane 6), HT-29 (Lane 7) and tissue extract of Mouse Testis (Lane 8). The blot was probed with Anti-hnRNP K Polyclonal Antibody (Product # PA5-27522, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 55 kDa band corresponding to hnRNP was observed across the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of hnRNP K was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a hnRNP K Polyclonal Antibody (Product # PA5-27522) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using hnRNP K Polyclonal Antibody (Product # PA5-27522). Sample (50 µg of whole cell lysate). Lane A: Mouse brain. 10% SDS PAGE. hnRNP K Polyclonal Antibody (Product # PA5-27522) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of hnRNP K was achieved by transfecting HeLa with hnRNP K specific siRNAs (Silencer® select Product # s6737, s6738). Western blot analysis (Fig. a) was performed using whole cell extracts from the hnRNP K knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with hnRNP K Polyclonal Antibody (Product # PA5-27522, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to hnRNP K.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of hnRNP K was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with hnRNP K Polyclonal Antibody (Product # PA5-27522) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of hnRNP K was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: hnRNP K Polyclonal Antibody (Product # PA5 27522) diluted at 1:500. Red: phalloidin, a cytoskeleton marker.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- hnRNP K Polyclonal Antibody detects hnRNP K protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded mouse brain. hnRNP K stained by hnRNP K Polyclonal Antibody (Product # PA5-27522) diluted at 1:1,000. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- hnRNP K Polyclonal Antibody detects hnRNP K protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded mouse testis. hnRNP K stained by hnRNP K Polyclonal Antibody (Product # PA5-27522) diluted at 1:1,000. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- hnRNP K Polyclonal Antibody detects hnRNP K protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded rat heart. hnRNP K stained by hnRNP K Polyclonal Antibody (Product # PA5-27522) diluted at 1:1,000. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- hnRNP K Polyclonal Antibody detects hnRNP K protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded rat kidney. hnRNP K stained by hnRNP K Polyclonal Antibody (Product # PA5-27522) diluted at 1:1,000. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- hnRNP K Polyclonal Antibody detects hnRNP K protein at nucleus on mouse colon by immunohistochemical analysis. Sample: Paraffin-embedded mouse colon. HnRNP K Polyclonal Antibody (Product # PA5-27522) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- hnRNP K Polyclonal Antibody detects hnRNP K protein at nucleus on mouse colon by immunohistochemical analysis. Sample: Paraffin-embedded mouse colon. HnRNP K Polyclonal Antibody (Product # PA5-27522) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- hnRNP K Polyclonal Antibody detects hnRNP K protein at nucleus on rat brain stem by immunohistochemical analysis. Sample: Paraffin-embedded rat brain stem. HnRNP K Polyclonal Antibody (Product # PA5-27522) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded H1299 xenograft, using hnRNP K (Product # PA5-27522) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Detection of binding of endogenous hnRNP K protein to specific RNA using Anti-hnRNP K Antibody: RNA Immunoprecipitation (RIP) was performed using Anti-hnRNP K Rabbit Polyclonal Antibody (Product # PA5-27522, 5 ug) on clarified whole cell lysate from 4 million HCT 116 cells. Normal Rabbit IgG was used as a negative IP control. Immunoprecipitated RNA was purified by RiboPure™ RNA Purification Kit (Product # AM1924) and analyzed by RT-PCR using the Power SYBR® Green RNA-to-CT™ 1-Step Kit (Product # 4389986) with primer pairs over ZFP36 and EGR1 (positive) and 18S rRNA (negative). Data is presented as the fraction of immunoprecipitated RNA (%IP) normalized to the total amount of RNA used for immunoprecipitation.
