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FCM (Flow Cytometry) (Figure 10. Flow Cytometry analysis of JK cells using anti-KLHL12 antibody (AAA19601).Overlay histogram showing JK cells stained with AAA19601 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-KLHL12 Antibody (AAA19601, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Rabbit KLHL12 Polyclonal Antibody | anti-KLHL12 antibody

Anti-KLHL12 Antibody Picoband

Gene Names
KLHL12; DKIR; C3IP1
Reactivity
Human, Mouse, Rat
Applications
ELISA, Flow Cytometry, Functional Assay, Immunofluorescence, Immunohistochemistry, Immunocytochemistry, Western Blot
Purity
Immunogen affinity purified.
Synonyms
KLHL12; Polyclonal Antibody; Anti-KLHL12 Antibody Picoband; anti-KLHL12 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Concentration
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml. (varies by lot)
Applicable Applications for anti-KLHL12 antibody
ELISA (EIA), Flow Cytometry (FC/FACS), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Western Blot (WB)
Application Notes
WB: 0.1-0.25 ug/ml, Human
IHC-P: 2-5 ug/ml, Human, Mouse, Rat
ICC/IF: 5 ug/ml, Human
FC/FACS: 1-3 ug/1x10^6 cells, Human
Direct ELISA: 0.1-0.5 ug/ml, Human
Tested Species: In-house tested species with positive results.
Enhanced Chemiluminescent Kit with anti-Rabbit IgG for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit for IHC(P) and ICC.
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml.
Immunogen
E Coli-derived human KLHL12 recombinant protein (Position: R27-I331).
Preparation and Storage
Store at -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months. Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 10. Flow Cytometry analysis of JK cells using anti-KLHL12 antibody (AAA19601).Overlay histogram showing JK cells stained with AAA19601 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-KLHL12 Antibody (AAA19601, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

FCM (Flow Cytometry) (Figure 10. Flow Cytometry analysis of JK cells using anti-KLHL12 antibody (AAA19601).Overlay histogram showing JK cells stained with AAA19601 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-KLHL12 Antibody (AAA19601, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IF (Immunofluorescence)

(Figure 9. IF analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in an immunocytochemical section of T47D cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/mL rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

IF (Immunofluorescence) (Figure 9. IF analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in an immunocytochemical section of T47D cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/mL rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

IHC (Immunohistochemistry)

(Figure 8. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of rat lymph node tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 8. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of rat lymph node tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 7. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 7. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistchemistry)

(Figure 6. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistchemistry) (Figure 6. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 5. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of human thyroid papillary carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 5. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of human thyroid papillary carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 4. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 3. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 3. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 2. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of human hashimoto thyroiditis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 2. IHC analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).KLHL12 was detected in a paraffin-embedded section of human hashimoto thyroiditis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-KLHL12 Antibody (AAA19601) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.Lane 1: human PC-3 whole cell lysates,Lane 2: human HEK293 whole cell lysates,Lane 3: human Hela whole cell lysates,Lane 4: human Jurkat whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-KLHL12 antigen affinity purified polyclonal antibody (#AAA19601) at 0.25 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for KLHL12 at approximately 63 kDa. The expected band size for KLHL12 is at 63 kDa.)

WB (Western Blot) (Figure 1. Western blot analysis of KLHL12 using anti-KLHL12 antibody (AAA19601).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.Lane 1: human PC-3 whole cell lysates,Lane 2: human HEK293 whole cell lysates,Lane 3: human Hela whole cell lysates,Lane 4: human Jurkat whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-KLHL12 antigen affinity purified polyclonal antibody (#AAA19601) at 0.25 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for KLHL12 at approximately 63 kDa. The expected band size for KLHL12 is at 63 kDa.)
Related Product Information for anti-KLHL12 antibody
Kelch-like protein 12 is a protein that in humans is encoded by the KLHL12 gene. This gene encodes a member of the KLHL (Kelch-like) family of proteins. This protein has been identified as an autoantigen in the autoimmune disease Sjogren's syndrome and as a potential biomarker in primary biliary cirrhosis. This protein may act as a substrate adaptor of the Cullin-3 ubiquitin ligase complex to promote substrate-specific ubiquitylation. Ubiquitylation by this complex has been shown to regulate the Wnt signaling pathway as well as COPII vesicle coat size. A pseudogene has been identified on chromosome 22. Alternative splicing results in multiple transcript variants.
Product Categories/Family for anti-KLHL12 antibody

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
UniProt Accession #
Molecular Weight
568
NCBI Official Full Name
Kelch-like protein 12
NCBI Official Synonym Full Names
kelch-like family member 12
NCBI Official Symbol
KLHL12
NCBI Official Synonym Symbols
DKIR; C3IP1
NCBI Protein Information
kelch-like protein 12; DKIR homolog; kelch-like protein C3IP1; CUL3-interacting protein 1
UniProt Protein Name
Kelch-like protein 12
UniProt Gene Name
KLHL12
UniProt Synonym Gene Names
C3IP1; hDKIR
UniProt Entry Name
KLH12_HUMAN

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Product Notes

The KLHL12 klhl12 (Catalog #AAA19601) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-KLHL12 Antibody Picoband reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's KLHL12 can be used in a range of immunoassay formats including, but not limited to, ELISA (EIA), Flow Cytometry (FC/FACS), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Western Blot (WB). WB: 0.1-0.25 ug/ml, Human IHC-P: 2-5 ug/ml, Human, Mouse, Rat ICC/IF: 5 ug/ml, Human FC/FACS: 1-3 ug/1x10^6 cells, Human Direct ELISA: 0.1-0.5 ug/ml, Human Tested Species: In-house tested species with positive results. Enhanced Chemiluminescent Kit with anti-Rabbit IgG for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit for IHC(P) and ICC. Researchers should empirically determine the suitability of the KLHL12 klhl12 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "KLHL12, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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